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151 results on '"Stabilité Génétique et Oncogenèse (UMR 8200)"'

1. NOX2-dependent ATM kinase activation dictates pro-inflammatory macrophage phenotype and improves effectiveness to radiation therapy

Author

Laurent Voisin, Céline Leteur, Fabien Milliat, Eric Deutsch, Qiuji Wu, Eric Solary, Zeinaf Muradova, Haithem Dakhli, Audrey Paoletti, Awatef Allouch, Jean-Luc Perfettini, Filippo Rosselli, Frédéric Law, Mélanie Gauthier, David M. Ojcius, Maxime Thoreau, Elodie Mintet, Nazanine Modjtahedi, Olivier Caron, Isabelle Martins, Céline Mirjolet, Radiothérapie moléculaire (UMR 1030), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Gustave Roussy (IGR)-Université Paris-Sud - Paris 11 (UP11), Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon] (UNICANCER/CRLCC-CGFL), UNICANCER, Institut Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de Recherche en Informatique, Signal et Automatique de Lille - UMR 9189 (CRIStAL), Centrale Lille-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Institut de radioprotection et de sûreté nucléaire [Fontenay-aux-Roses] (IRSN), Ministère de l'économie, de l'industrie et de l'emploi-Ministère de la Défense-Ministère de la santé-Ministère de l'Enseignement Supérieur et de la Recherche Scientifique-Ministère de l'écologie de l'Energie, du Développement durable et de l'Aménagement du territoire, University of California [Merced], University of California, Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Apoptose, cancer et immunité (U848), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Radiothérapie moléculaire [UMR 1030], Centre Régional de Lutte contre le cancer Georges-François Leclerc [Dijon] [UNICANCER/CRLCC-CGFL], Institut Jacques Monod [IJM (UMR_7592)], Institut Cochin [IC UM3 (UMR 8104 / U1016)], Centre de Recherche en Informatique, Signal et Automatique de Lille - UMR 9189 [CRIStAL], University of California [Merced] [UC Merced], Stabilité Génétique et Oncogenèse [UMR 8200], Apoptose, cancer et immunité [U848], Institut Gustave Roussy (IGR), Systerel, Institut de Radioprotection et de Sûreté Nucléaire (IRSN), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Onco-génétique, Département de médecine oncologique [Gustave Roussy], Institut Gustave Roussy (IGR)-Institut Gustave Roussy (IGR), Laboratoire de Radiopathologie, Génomes et cancer (GC (FRE2939)), Hématopoïèse normale et pathologique, This work was supported by funds from Agence Nationale de la Recherche (ANR-10-IBHU-0001, ANR-10-LABX33 and ANR-11-IDEX-003-01), Electricité de France, Fondation Gustave Roussy, Institut National du Cancer (INCA 9414), NATIXIS, SIDACTION and the French National Agency for Research on AIDS and viral Hepatitis (ANRSH) (to J-LP.), Electricité de France and Fondation Gustave Roussy (to ED). QW is recipient of PhD fellowship of China Scholarship Council. AP and AA are, respectively, recipient of PhD fellowship and post-doc fellowship from Agence Nationale de Recherche sur le Sida et sur les Hépatites (ANRSH). LV and FL are recipient of PhD fellowships from Fondation pour la Recherche Médicale and CIFRE. HD, EM and MT are supported by the Laboratory of Excellence LERMIT with a grant from ANR (ANR-10-LABX-33) under the program ‘Investissements d'Avenir’ ANR-11-IDEX-0003-01. IM is funded by INCA (INCA-DGOS-INSERM 6043). CM work was supported by the ‘Cancéropôle Grand Est’, and the ‘Conseils Régionaux de Bourgogne, de Franche Comté et de Lorraine’., We gratefully acknowledge S Solier, Y Lecluse and S Salome-Desnoulez for technical support., Centre Régional de Lutte contre le cancer - Centre Georges-François Leclerc (CRLCC - CGFL), Centre de Recherche en Informatique, Signal et Automatique de Lille (CRIStAL) - UMR 9189 (CRIStAL), Centre National de la Recherche Scientifique (CNRS)-Université de Lille-Ecole Centrale de Lille, and Ministère de l'écologie de l'Energie, du Développement durable et de l'Aménagement du territoire-Ministère de la santé-Ministère de la Défense-Ministère de l'Enseignement Supérieur et de la Recherche Scientifique-Ministère de l'économie, de l'industrie et de l'emploi

Subjects
0301 basic medicine, [SDV]Life Sciences [q-bio], Ataxia Telangiectasia Mutated Proteins, Neurodegenerative, Medical and Health Sciences, Mice, 0302 clinical medicine, Macrophage, Phosphorylation, ComputingMilieux_MISCELLANEOUS, Cancer, Microscopy, NADPH oxidase, biology, Kinase, Biological Sciences, Flow Cytometry, 3. Good health, Cell biology, 030220 oncology & carcinogenesis, Signal transduction, Signal Transduction, Biochemistry & Molecular Biology, Fluorescence, Cell Line, 03 medical and health sciences, Interferon-gamma, Ataxia Telangiectasia, Rare Diseases, Genetics, Animals, Humans, Molecular Biology, Protein Processing, Original Paper, Macrophages, Post-Translational, Cell Biology, Macrophage Activation, 030104 developmental biology, RAW 264.7 Cells, Microscopy, Fluorescence, Apoptosis, biology.protein, Cancer research, Protein Processing, Post-Translational, IRF5, Interferon regulatory factors
Abstract

International audience; Although tumor-associated macrophages have been extensively studied in the control of response to radiotherapy, the molecular mechanisms involved in the ionizing radiation-mediated activation of macrophages remain elusive. Here we show that ionizing radiation induces the expression of interferon regulatory factor 5 (IRF5) promoting thus macrophage activation toward a pro-inflammatory phenotype. We reveal that the activation of the ataxia telangiectasia mutated (ATM) kinase is required for ionizing radiation-elicited macrophage activation, but also for macrophage reprogramming after treatments with γ-interferon, lipopolysaccharide or chemotherapeutic agent (such as cisplatin), underscoring the fact that the kinase ATM plays a central role during macrophage phenotypic switching toward a pro-inflammatory phenotype through the regulation of mRNA level and post-translational modifications of IRF5. We further demonstrate that NADPH oxidase 2 (NOX2)-dependent ROS production is upstream to ATM activation and is essential during this process. We also report that the inhibition of any component of this signaling pathway (NOX2, ROS and ATM) impairs pro-inflammatory activation of macrophages and predicts a poor tumor response to preoperative radiotherapy in locally advanced rectal cancer. Altogether, our results identify a novel signaling pathway involved in macrophage activation that may enhance the effectiveness of radiotherapy through the reprogramming of tumor-infiltrating macrophages.

Published
2017

2. A single aspartate mutation in the conserved catalytic site of Rev3L generates a hypomorphic phenotype in vivo and in vitro

Author

Sébastien Storck, Benoit Palancade, Annie De Smet, Frédéric Delbos, J.C. Weill, Said Aoufouchi, Christine E. Canman, Rémi Fritzen, C.A. Reynaud, Institut Necker Enfants-Malades (INEM - UM 111 (UMR 8253 / U1151)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université Sorbonne Paris Cité (USPC), Institut Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Department of Pharmacology [Ann Arbor], University of Michigan [Ann Arbor], University of Michigan System-University of Michigan System, Réparation de l’ADN (CNRS UMR 8200), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Ligue Nationale contre le Cancer ('Equipe labellisée'), Fondation Princesse Grace, ANR-06-BLAN-0182,MUTIG,Hypermutation des gènes des immunoglobulines: étude du rôle des ADN polymérases translésionnelles par transfert de cellules souches hématopoiétiques modifiées(2006), Institut Necker Enfants-Malades (INEM) ( INEM - UM 111 (UMR 8253 / U1151) ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Université Sorbonne Paris Cité ( USPC ), Institut Jacques Monod ( IJM ), Université Paris Diderot - Paris 7 ( UPD7 ) -Centre National de la Recherche Scientifique ( CNRS ), Department of Pharmacology, Michigan, University of Michigan, Réparation de l’ADN ( CNRS UMR 8200 ), Stabilité Génétique et Oncogenèse ( UMR 8200 ), Université Paris-Sud - Paris 11 ( UP11 ) -Institut Gustave Roussy ( IGR ) -Centre National de la Recherche Scientifique ( CNRS ) -Université Paris-Sud - Paris 11 ( UP11 ) -Institut Gustave Roussy ( IGR ) -Centre National de la Recherche Scientifique ( CNRS ), and ANR-06-BLAN-0182,MUTIG,Hypermutation des gènes des immunoglobulines: étude du rôle des ADN polymérases translésionnelles par transfert de cellules souches hématopoiétiques modifiées ( 2006 )

Subjects
0301 basic medicine, Immunoglobulin gene, Base pair, DNA polymerase, DNA damage, Cell Survival, Ultraviolet Rays, [ SDV.TOX ] Life Sciences [q-bio]/Toxicology, Mutant, Somatic hypermutation, Gene Expression, Immunoglobulins, Mice, Transgenic, DNA-Directed DNA Polymerase, Biochemistry, Cell Line, 03 medical and health sciences, Mice, REV3L, Rev3l, Catalytic Domain, Animals, Humans, Amino Acid Sequence, Molecular Biology, Conserved Sequence, DNA polymerase zeta, Phenocopy, Aspartic Acid, [SDV.GEN]Life Sciences [q-bio]/Genetics, Alanine, biology, Cell Biology, Fibroblasts, Embryo, Mammalian, Translesion, Molecular biology, DNA-Binding Proteins, 030104 developmental biology, HEK293 Cells, Phenotype, Amino Acid Substitution, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Mutation, biology.protein, [ SDV.GEN ] Life Sciences [q-bio]/Genetics
Abstract

International audience; Rev3, the catalytic subunit of yeast DNA polymerase ζ, is required for UV resistance and UV-induced mutagenesis, while its mammalian ortholog, REV3L, plays further vital roles in cell proliferation and embryonic development. To assess the contribution of REV3L catalytic activity to its in vivo function, we generated mutant mouse strains in which one or two Ala residues were substituted to the Asp of the invariant catalytic YGDTDS motif. The simultaneous mutation of both Asp (ATA) phenocopies the Rev3l knockout, which proves that the catalytic activity is mandatory for the vital functions of Rev3L, as reported recently. Surprisingly, although the mutation of the first Asp severely impairs the enzymatic activity of other B-family DNA polymerases, the corresponding mutation of Rev3 (ATD) is hypomorphic in yeast and mouse, as it does not affect viability and proliferation and moderately impacts UVC-induced cell death and mutagenesis. Interestingly, Rev3l hypomorphic mutant mice display a distinct, albeit modest, alteration of the immunoglobulin gene mutation spectrum at G-C base pairs, further documenting its role in this process.

Published
2016

3. Analysis of the role of autophagy inhibition by two complementary human cytomegalovirus BECN1/Beclin 1-binding proteins

Author

Rebekka Brost, Lina Mouna, Adam P. Geballe, Wolfram Brune, Audrey Esclatine, Isabelle Beau, Dorine Bonte, Laura Ruth Delgui, Larbi Amazit, Eva Hernandez, Virulence et Latence des Herpesvirus ( HERPES ), Département Virologie ( Dpt Viro ), Institut de Biologie Intégrative de la Cellule ( I2BC ), Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Sud - Paris 11 ( UP11 ) -Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Sud - Paris 11 ( UP11 ) -Institut de Biologie Intégrative de la Cellule ( I2BC ), Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Sud - Paris 11 ( UP11 ) -Université Paris-Saclay-Centre National de la Recherche Scientifique ( CNRS ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Sud - Paris 11 ( UP11 ), Stabilité Génétique et Oncogenèse ( UMR 8200 ), Université Paris-Sud - Paris 11 ( UP11 ) -Institut Gustave Roussy ( IGR ) -Centre National de la Recherche Scientifique ( CNRS ), Leibniz Institute for Experimental Virology, Heinrich Pette Institute, Signalisation Hormonale, Physiopathologie Endocrinienne et Métabolique, Université Paris-Sud - Paris 11 ( UP11 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -AP-HP Hôpital Bicêtre (Le Kremlin-Bicêtre), Fred Hutchinson Cancer Research Center and University of Washington, Seattle, WA, USA, Fred Hutchinson Cancer Research Center [Seattle] ( FHCRC ), Virulence et Latence des Herpesvirus (HERPES), Département Virologie (Dpt Viro), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), AP-HP Hôpital Bicêtre (Le Kremlin-Bicêtre)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Sud - Paris 11 (UP11), University of Washington [Seattle], and Université Paris-Sud - Paris 11 (UP11)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AP-HP Hôpital Bicêtre (Le Kremlin-Bicêtre)

Subjects
0301 basic medicine, Male, Programmed cell death, endocrine system, Irs1, [SDV]Life Sciences [q-bio], Otras Ciencias Biológicas, viruses, Becn1, Cytomegalovirus, Biology, BAG3, Virus, Ciencias Biológicas, purl.org/becyt/ford/1 [https], 03 medical and health sciences, Viral Proteins, eIF-2 Kinase, Protein Domains, Eif2ak2/Pkr, Autophagy, Humans, purl.org/becyt/ford/1.6 [https], Molecular Biology, Recombination, Genetic, EIF-2 kinase, [ SDV ] Life Sciences [q-bio], Cell Biology, BECN1, Protein kinase R, Basic Research Paper, 3. Good health, Cell biology, 030104 developmental biology, Viral replication, Cytomegalovirus Infections, Mutation, Cancer research, biology.protein, Beclin-1, Trs1, CIENCIAS NATURALES Y EXACTAS, HeLa Cells, Protein Binding
Abstract

Autophagy is activated early after human cytomegalovirus (HCMV) infection, but lateron the virus blocks autophagy. Here we characterized 2 HCMV proteins, TRS1 andIRS1, which inhibit autophagy during infection. Expression of either TRS1 or IRS1 wasable to block autophagy in different cell lines, independently of the EIF2S1 kinase,EIF2AK2/PKR. Instead, TRS1 and IRS1 interacted with the autophagy proteinBECN1/Beclin 1. We mapped the BECN1-binding domain (BBD) of IRS1 and TRS1 andfound it to be essential for autophagy inhibition. Mutant viruses that express only IRS1or TRS1 partially controlled autophagy, whereas a double mutant virus expressingneither protein stimulated autophagy. A mutant virus that did not express IRS1 andexpressed a truncated form of TRS1 in which the BBD was deleted, failed to controlautophagy. However, this mutant virus had similar replication kinetics as wild-type virus,suggesting that autophagy inhibition is not critical for viral replication. In fact, usingpharmacological modulators of autophagy and inhibition of autophagy by shRNAknockdown, we discovered that stimulating autophagy enhanced viral replication.Conversely, inhibiting autophagy decreased HCMV infection. Thus, our resultsdemonstrate a new proviral role of autophagy for a DNA virus Fil: Mouna, Lina. Université Paris Sud; Francia Fil: Hernandez, Eva. Université Paris Sud; Francia Fil: Bonte, Dorine. Centre National de la Recherche Scientifique; Francia Fil: Brost, Rebekka. Heinrich Pette Institute; Alemania Fil: Amazit, Larbi. Université Paris Sud; Francia Fil: Delgui, Laura Ruth. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina Fil: Brune, Wolfram. Heinrich Pette Institute; Alemania Fil: Geballe, Adam P.. University of Washington; Estados Unidos Fil: Beau, Isabelle. Université Paris Sud; Francia Fil: Esclatine, Audrey. Université Paris Sud; Francia

Published
2015

4. Distinct deregulation of the hypoxia inducible factor by PHD2 mutants identified in germline DNA of patients with polycythemia

Author

Romain Carcenac, Michel Leporrier, Stéphane Richard, Michel Barrois, Jean-Jacques Kiladjian, Jacques Pouysségur, André Baruchel, Nathalie M. Mazure, David Hoogewijs, Roland H. Wenger, Anne-Paule Gimenez-Roqueplo, Charline Ladroue, Brigitte Bressac-de Paillerets, Nicole Casadevall, Jean Feunteun, Betty Gardie, Sophie Gad, Fadi Fakhoury, Olivier Hermine, Federica Storti, Bernardo, Elizabeth, Cytokines et Immunologie des Tumeurs Humaines (U753), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Zürich Center for Integrative Human Physiology (ZIHP), Universität Zürich [Zürich] = University of Zurich (UZH)-Institute of Physiology, Service de Génétique [Villejuif], Institut Gustave Roussy (IGR), Laboratoire de Recherche en Imagerie : Méthodes d'imagerie des Échanges transcapillaires (LRI - EA4062), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service de Génétique [CHU HEGP], Hôpital Européen Georges Pompidou [APHP] (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO), Laboratoire d'Hématologie Biologique [CHU Caen], Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN), Hematopoïese et cellules souches normales et pathologiques (U790), Service d'immuno-hématologie pédiatrique [CHU Necker], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Centre d'Investigations Cliniques, Hopital Saint-Louis [AP-HP] (AP-HP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Service d'hématologie greffe [Saint-Louis], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Diderot - Paris 7 (UPD7)-Groupe Hospitalier Saint Louis - Lariboisière - Fernand Widal [Paris], Département de Néphrologie [CHU Necker], Variabilité Génétique et Maladies Humaines, Institut Universitaire d'Hématologie (IUH), Université Paris Diderot - Paris 7 (UPD7)-Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Institut de signalisation, biologie du développement et cancer (ISBDC), Université Nice Sophia Antipolis (1965 - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Centre National de la Recherche Scientifique (CNRS)-Université Côte d'Azur (UCA), Centre Expert National Cancers Rares INCa 'PREDIR' and Réseau National INCa [AP-HP Hôpital Bicêtre, Paris], Service d'urologie, Université Paris-Sud - Paris 11 (UP11)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Bicêtre-Université Paris-Sud - Paris 11 (UP11)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Bicêtre, This work was supported by grants from the 'Association pour la Recherche sur le Cancer' (ARC), the French National Cancer Institute ('INCa', PNES Rein, and Réseau National Prédispositions héréditaires au cancer du rein) and the French Ligue Nationale contre le Cancer (Comités du Cher et de l’Indre) as well as by the HypoxiaNet COST Action TD0901. DH is supported by a postdoctoral Marie Curie IEF Fellowship from the European Commission and the Sassella Stiftung. DH and RHW are supported by grants from the State Secretariat of Education and Research (C10.0106) and from the NCCR Kidney.CH, funded by the SNF., Cytokines et Immunologie des Tumeurs Humaines ( U753 ), Université Paris-Sud - Paris 11 ( UP11 ) -Institut Gustave Roussy ( IGR ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Zürich Center for Integrative Human Physiology ( ZIHP ), University of Zürich [Zürich] ( UZH ) -Institute of Physiology, Institut Gustave Roussy ( IGR ), Laboratoire de Recherche en Imagerie : Méthodes d'imagerie des Échanges transcapillaires ( LRI - EA4062 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Service de Génétique [AP-HP Hôpital Européen Georges Pompidou, Paris], Hôpital Européen Georges Pompidou [APHP] ( HEGP ), Service d'Hématologie Clinique, CHU Caen-Hôpital Clémenceau, Hematopoïese et cellules souches normales et pathologiques ( U790 ), Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Necker - Enfants Malades [AP-HP], CHU Saint Louis [APHP], Assistance publique - Hôpitaux de Paris (AP-HP)-Université Paris Diderot - Paris 7 ( UPD7 ) -Groupe Hospitalier Saint Louis - Lariboisière - Fernand Widal [Paris], Université Paris Diderot - Paris 7 ( UPD7 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Stabilité Génétique et Oncogenèse ( UMR 8200 ), Université Paris-Sud - Paris 11 ( UP11 ) -Institut Gustave Roussy ( IGR ) -Centre National de la Recherche Scientifique ( CNRS ), Institut de signalisation, biologie du développement et cancer ( ISBDC ), Université Nice Sophia Antipolis ( UNS ), Université Côte d'Azur ( UCA ) -Université Côte d'Azur ( UCA ) -Centre National de la Recherche Scientifique ( CNRS ), Université Paris-Sud - Paris 11 ( UP11 ) -Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Bicêtre-Université Paris-Sud - Paris 11 ( UP11 ) -Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Bicêtre, Centre National de la Recherche Scientifique (CNRS)-Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Université Côte d'Azur (UCA), and Hôpital Bicêtre-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris-Sud - Paris 11 (UP11)-Hôpital Bicêtre-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris-Sud - Paris 11 (UP11)

Subjects
Adult, Male, hypoxia inducible factor, Adolescent, Tumor suppressor gene, Mutant, Procollagen-Proline Dioxygenase, [SDV.CAN]Life Sciences [q-bio]/Cancer, Polycythemia, Biology, medicine.disease_cause, Germline, Hypoxia-Inducible Factor-Proline Dioxygenases, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, Hydroxylation, Young Adult, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Germline mutation, [SDV.CAN] Life Sciences [q-bio]/Cancer, erythrocytosis, medicine, Humans, PHD2, Cells, Cultured, Germ-Line Mutation, hypoxia-inducible transcription factor, 030304 developmental biology, 0303 health sciences, Base Sequence, Hydrolysis, Original Articles, Hematology, Middle Aged, Hypoxia (medical), Hypoxia-Inducible Factor 1, alpha Subunit, Molecular biology, 3. Good health, HEK293 Cells, chemistry, Hypoxia-inducible factors, 030220 oncology & carcinogenesis, Female, Mutant Proteins, medicine.symptom, Carcinogenesis
Abstract

International audience; BACKGROUND: Congenital secondary erythrocytoses are due to deregulation of hypoxia inducible factor resulting in overproduction of erythropoietin. The most common germline mutation identified in the hypoxia signaling pathway is the Arginine 200-Tryptophan mutant of the von Hippel-Lindau tumor suppressor gene, resulting in Chuvash polycythemia. This mutant displays a weak deficiency in hypoxia inducible factor α regulation and does not promote tumorigenesis. Other von Hippel-Lindau mutants with more deleterious effects are responsible for von Hippel-Lindau disease, which is characterized by the development of multiple tumors. Recently, a few mutations in gene for the prolyl hydroxylase domain 2 protein (PHD2) have been reported in cases of congenital erythrocytosis not associated with tumor formation with the exception of one patient with a recurrent extra-adrenal paraganglioma.DESIGN AND METHODS: Five PHD2 variants, four of which were novel, were identified in patients with erythrocytosis. These PHD2 variants were functionally analyzed and compared with the PHD2 mutant previously identified in a patient with polycythemia and paraganglioma. The capacity of PHD2 to regulate the activity, stability and hydroxylation of hypoxia inducible factor α was assessed using hypoxia-inducible reporter gene, one-hybrid and in vitro hydroxylation assays, respectively.RESULTS: This functional comparative study showed that two categories of PHD2 mutants could be distinguished: one category with a weak deficiency in hypoxia inducible factor α regulation and a second one with a deleterious effect; the mutant implicated in tumor occurrence belongs to the second category.CONCLUSIONS: As observed with germline von Hippel-Lindau mutations, there are functional differences between the PHD2 mutants with regards to hypoxia inducible factor regulation. PHD2 mutation carriers do, therefore, need careful medical follow-up, since some mutations must be considered as potential candidates for tumor predisposition.

Published
2011

5. Genetic evidence of a precisely tuned dysregulation in the hypoxia signaling pathway during oncogenesis

Author

Sophie Couvé, Hélène Le Jeune, David R. Mole, Karène Mahtouk, Gregory Nuel, Peter J. Ratcliffe, Anne Paule Gimenez-Roqueplo, Justine Guegan, Françoise Lasne, William G. Kaelin, Nathalie M. Mazure, Charline Ladroue, Jean Christophe Pagès, William Y. Kim, Patrick R. Benusiglio, Luba Tchertanov, Stéphane Richard, Brigitte Bressac-de Paillerets, Marion Le Gentil, Vladimir Lazar, Elodie Laine, Philippe Dessen, Betty Gardie, Christine Collin, Jean Feunteun, Sophie Gad, Bernard Lecomte, Institut National du Cancer [Boulogne Billancourt] (INC), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL), Cytokines et Immunologie des Tumeurs Humaines (U753), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Biologie Computationnelle et Quantitative = Laboratory of Computational and Quantitative Biology (LCQB), Institut de Biologie Paris Seine (IBPS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Plateforme de Génomique [Gustave Roussy], Analyse moléculaire, modélisation et imagerie de la maladie cancéreuse (AMMICa), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Mathématiques Appliquées Paris 5 (MAP5 - UMR 8145), Université Paris Descartes - Paris 5 (UPD5)-Institut National des Sciences Mathématiques et de leurs Interactions (INSMI)-Centre National de la Recherche Scientifique (CNRS), Lineberger Comprehensive Cancer Center (UNC Lineberger), University of North Carolina [Chapel Hill] (UNC), University of North Carolina System (UNC)-University of North Carolina System (UNC), Médecine générale, Virus, pseudo-virus: Morphogénèse et Antigénicité, Université de Tours-EA3856, Agence Française de Lutte contre le Dopage (AFLD), Institut Gustave Roussy (IGR), Onco-génétique, Département de médecine oncologique [Gustave Roussy], Institut Gustave Roussy (IGR)-Institut Gustave Roussy (IGR), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Angiogenesis Imaging UMR970, Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Recherche sur le Cancer et le Vieillissement (IRCAN), Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Côte d'Azur (UCA), Henry Wellcome Building for Molecular Physiology, University of Oxford [Oxford], Howard Hughes Medical Institute (HHMI), Génétique Oncologique EPHE, Faculté de Médecine Paris-Sud, Le Kremlin-Bicêtre, Centre de Recherche en Cancérologie Nantes-Angers (CRCNA), Centre Hospitalier Universitaire d'Angers (CHU Angers), PRES Université Nantes Angers Le Mans (UNAM)-PRES Université Nantes Angers Le Mans (UNAM)-Hôtel-Dieu de Nantes-Institut National de la Santé et de la Recherche Médicale (INSERM)-Hôpital Laennec-Centre National de la Recherche Scientifique (CNRS)-Faculté de Médecine d'Angers-Centre hospitalier universitaire de Nantes (CHU Nantes), This work was supported by grants from the Ligue Nationale contre le Cancer (Comites du Cher et de l'Indre, de la Loire Atlantique et des Cotes d'Armor), the French NCI (INCa, PNES 'Kidney cancer'), the 'Association pour la Recherche sur le Cancer' (ARC), the 'Association VHL France,' the Region Pays de la Loire, as well as by the HypoxiaNet COST Action TD0901. S. Couve was supported by postdoctoral fellowships from the Fondation 'Cancer, Aidez la recherche!' and the 'Fondation Gustave Roussy.'The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact., Bernardo, Elizabeth, Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Biologie Paris Seine (IBPS), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Université de Tours (UT)-EA3856, Université Nice Sophia Antipolis (1965 - 2019) (UNS), University of Oxford, Institut National du Cancer [Boulogne Billancourt] ( INC ), École pratique des hautes études ( EPHE ), Cytokines et Immunologie des Tumeurs Humaines ( U753 ), Université Paris-Sud - Paris 11 ( UP11 ) -Institut Gustave Roussy ( IGR ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Biologie Computationnelle et Quantitative = Laboratory of Computational and Quantitative Biology ( LCQB ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Centre National de la Recherche Scientifique ( CNRS ), Plateforme de génomique, Analyse moléculaire, modélisation et imagerie de la maladie cancéreuse ( AMMICa ), Université Paris-Sud - Paris 11 ( UP11 ) -Institut Gustave Roussy ( IGR ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ) -Université Paris-Sud - Paris 11 ( UP11 ) -Institut Gustave Roussy ( IGR ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Mathématiques Appliquées à Paris 5 ( MAP5 - UMR 8145 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National des Sciences Mathématiques et de leurs Interactions-Centre National de la Recherche Scientifique ( CNRS ), Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill, Agence Française de Lutte contre le Dopage ( AFLD ), Institut Gustave Roussy ( IGR ), Service de Génétique, Institut de cancérologie Gustave Roussy, Villejuif, France, Stabilité Génétique et Oncogenèse ( UMR 8200 ), Université Paris-Sud - Paris 11 ( UP11 ) -Institut Gustave Roussy ( IGR ) -Centre National de la Recherche Scientifique ( CNRS ), Institut National de la Santé et de la Recherche Médicale ( INSERM ), Institut de Recherche sur le Cancer et le Vieillissement ( IRCAN ), Centre National de la Recherche Scientifique ( CNRS ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Nice Sophia Antipolis ( UNS ), Université Côte d'Azur ( UCA ) -Université Côte d'Azur ( UCA ), Howard Hugues Medical Institute, Centre de Recherche en Cancérologie / Nantes - Angers ( CRCNA ), CHU Angers-Centre hospitalier universitaire de Nantes ( CHU Nantes ) -Hôtel-Dieu de Nantes-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Hôpital Laennec-Centre National de la Recherche Scientifique ( CNRS ) -Faculté de Médecine d'Angers, École Pratique des Hautes Études (EPHE), and Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Cancer Research, Pathology, medicine.medical_specialty, endocrine system diseases, Carcinogenesis, Mutant, [SDV.CAN]Life Sciences [q-bio]/Cancer, Pheochromocytoma, Biology, Molecular Dynamics Simulation, medicine.disease_cause, Compound heterozygosity, urologic and male genital diseases, Polymorphism, Single Nucleotide, Article, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, Malignant transformation, 03 medical and health sciences, 0302 clinical medicine, Germline mutation, [SDV.CAN] Life Sciences [q-bio]/Cancer, medicine, Basic Helix-Loop-Helix Transcription Factors, Humans, Gene, neoplasms, Carcinoma, Renal Cell, 030304 developmental biology, 0303 health sciences, medicine.disease, Phenotype, female genital diseases and pregnancy complications, Kidney Neoplasms, 3. Good health, Oncology, Von Hippel-Lindau Tumor Suppressor Protein, 030220 oncology & carcinogenesis, Mutation, Cancer research, Signal Transduction
Abstract

The classic model of tumor suppression implies that malignant transformation requires full “two-hit” inactivation of a tumor-suppressor gene. However, more recent work in mice has led to the proposal of a “continuum” model that involves more fluid concepts such as gene dosage-sensitivity and tissue specificity. Mutations in the tumor-suppressor gene von Hippel-Lindau (VHL) are associated with a complex spectrum of conditions. Homozygotes or compound heterozygotes for the R200W germline mutation in VHL have Chuvash polycythemia, whereas heterozygous carriers are free of disease. Individuals with classic, heterozygous VHL mutations have VHL disease and are at high risk of multiple tumors (e.g., CNS hemangioblastomas, pheochromocytoma, and renal cell carcinoma). We report here an atypical family bearing two VHL gene mutations in cis (R200W and R161Q), together with phenotypic analysis, structural modeling, functional, and transcriptomic studies of these mutants in comparison with classical mutants involved in the different VHL phenotypes. We demonstrate that the complex pattern of disease manifestations observed in VHL syndrome is perfectly correlated with a gradient of VHL protein (pVHL) dysfunction in hypoxia signaling pathways. Thus, by studying naturally occurring familial mutations, our work validates in humans the “continuum” model of tumor suppression. Cancer Res; 74(22); 6554–64. ©2014 AACR.

Published
2014

6. In vivo inactivation of RAD51-mediated homologous recombination leads to premature aging, but not to tumorigenesis

Author

Gabriel Matos-Rodrigues, Vilma Barroca, Ali-Akbar Muhammad, Awatef Allouch, Stephane Koundrioukoff, Daniel Lewandowski, Emmanuelle Despras, Josée Guirouilh-Barbat, Lucien Frappart, Patricia Kannouche, Pauline Dupaigne, Eric Le Cam, Jean-Luc Perfettini, Paul-Henri Romeo, Michelle Debatisse, Maria Jasin, Gabriel Livera, Emmanuelle Martini, Bernard S. Lopez, Institut Cochin (IC UM3 (UMR 8104 / U1016)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Laboratoire de Développement des Gonades, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Institut de Radiobiologie Cellulaire et Moléculaire (IRCM), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Signalisation, noyaux et innovations en cancérologie (UMR8126), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Radiothérapie Moléculaire et Innovation Thérapeutique (RaMo-IT), Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay, Stabilité Génétique et Oncogenèse (UMR 8200), Leibniz Institute on Aging - Fritz Lipmann Institute (FLI), Leibniz Association, Radiothérapie moléculaire (UMR 1030), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Memorial Sloan Kettering Cancer Center (MSKCC), Stabilité génétique, cellules souches et radiations (SGCSR (U_1274 / UMR_E_008)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay-Université Paris Cité (UPCité), Intégrité du génome et cancers (IGC), École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Gustave Roussy (IGR)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Institut Gustave Roussy (IGR), and Memorial Sloane Kettering Cancer Center [New York]

Subjects
Genetics Homologous recombination, tumorigenesis, [SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics, mouse model, [SDV]Life Sciences [q-bio], aging, RAD51, [SDV.CAN]Life Sciences [q-bio]/Cancer, Biological Sciences
Abstract

Genetic instability is a hallmark of both cancer and aging. Homologous recombination (HR) is a prominent DNA repair pathway maintaining genomic integrity. Mutations in many HR genes lead to cancer predisposition. Paradoxically, the consequences of mutations in the pivotal HR player, RAD51, on cancer development remain puzzling. Moreover, in contrast with other HR genes, RAD51 mouse models are not available to experimentally address the role of RAD51 on aging and carcinogenesis, in vivo. Here, we engineered a mouse model with an inducible dominant negative form of RAD51 (SMRad51) that suppresses RAD51-mediated HR without stimulating alternative non-conservative repair pathways. We found that, in vivo expression of SMRad51 did not trigger tumorigenesis, but instead induced premature aging. We propose that these in vivo phenotypes result from the exhaustion of proliferating progenitors submitted to chronic endogenous replication stress resulting from RAD51-mediated HR suppression. Our data underline the importance of the RAD51 activity for progenitors homeostasis, preventing aging, and more generally for the balance between cancer and aging.

Published
2022

7. BRN2 is a non-canonical melanoma tumor-suppressor

Author

Pierre Sohier, Luis Sánchez-del-Campo, Nisamanee Charoenchon, Colin Kenny, Madeleine Le Coz, Colin R. Goding, Lionel Larue, Véronique Delmas, Valérie Petit, Alain Sarasin, Eiríkur Steingrímsson, Zackie Aktary, Jérémy H. Raymond, Dies Meijer, Franck Gesbert, Michael Hamm, Irwin Davidson, Robert A. Cornell, Laura Mosteo, Florian Rambow, Alfonso Bellacosa, Göran Jönsson, Marie Pouteaux, Martin Lauss, Signalisation normale et pathologique de l'embryon aux thérapies innovantes des cancers, Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Carver College of Medicine [Iowa City], University of Iowa [Iowa City], Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Mahidol University [Bangkok], Fox Chase Cancer Center, University of Oxford [Oxford], Lund University [Lund], University of Edinburgh, University of Iceland [Reykjavik], Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Signalisation, radiobiologie et cancer, Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Carver College of Medicine, University of Iowa, University of Oxford, and delmas, veronique

Subjects
0301 basic medicine, Neuroblastoma RAS viral oncogene homolog, Skin Neoplasms, Carcinogenesis, [SDV]Life Sciences [q-bio], General Physics and Astronomy, Haploinsufficiency, Cohort Studies, Mice, Phosphatidylinositol 3-Kinases, 0302 clinical medicine, CDKN2A, Genes, Tumor Suppressor, RNA, Small Interfering, Melanoma, Multidisciplinary, biology, Microphthalmia-associated transcription factor, Immunohistochemistry, Gene Expression Regulation, Neoplastic, Mechanisms of disease, Gene Knockdown Techniques, 030220 oncology & carcinogenesis, Disease Progression, [SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Proto-Oncogene Proteins B-raf, Chromatin Immunoprecipitation, DNA Copy Number Variations, Science, Mice, Transgenic, [SDV.CAN]Life Sciences [q-bio]/Cancer, Context (language use), Article, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, [SDV.CAN] Life Sciences [q-bio]/Cancer, Cell Line, Tumor, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], medicine, Animals, Humans, PTEN, Transcription factor, neoplasms, Cell Proliferation, Homeodomain Proteins, Microphthalmia-Associated Transcription Factor, POU domain, PTEN Phosphohydrolase, General Chemistry, Microarray Analysis, medicine.disease, Mice, Inbred C57BL, 030104 developmental biology, Mutation, POU Domain Factors, biology.protein, Cancer research, Gene expression
Abstract

While the major drivers of melanoma initiation, including activation of NRAS/BRAF and loss of PTEN or CDKN2A, have been identified, the role of key transcription factors that impose altered transcriptional states in response to deregulated signaling is not well understood. The POU domain transcription factor BRN2 is a key regulator of melanoma invasion, yet its role in melanoma initiation remains unknown. Here, in a BrafV600E PtenF/+ context, we show that BRN2 haplo-insufficiency promotes melanoma initiation and metastasis. However, metastatic colonization is less efficient in the absence of Brn2. Mechanistically, BRN2 directly induces PTEN expression and in consequence represses PI3K signaling. Moreover, MITF, a BRN2 target, represses PTEN transcription. Collectively, our results suggest that on a PTEN heterozygous background somatic deletion of one BRN2 allele and temporal regulation of the other allele elicits melanoma initiation and progression., The transcription factor BRN2 regulates melanoma migration and invasion, but its role during melanoma initiation is unclear. Here the authors show that BRN2 is a haplo-insufficient tumour suppressor that positively regulates PTEN expression and in the context of BRAF mutation and heterozygous PTEN, BRN2 loss promotes melanoma initiation and progression.

Published
2021

8. Insight into DNA substrate specificity of PARP1-catalysed DNA poly(ADP-ribosyl)ation

Author

Alexander A. Ishchenko, Bakhyt T. Matkarimov, Assel Kiribayeva, Bekbolat Khassenov, Elie Matta, Réparation de l’ADN (CNRS UMR 8200), Stabilité Génétique et Oncogenèse (UMR 8200), and Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, DNA damage, Poly (ADP-Ribose) Polymerase-1, lcsh:Medicine, Catalysis, Article, law.invention, Substrate Specificity, 03 medical and health sciences, chemistry.chemical_compound, Residue (chemistry), PARP1, law, Transferases, DNA-binding proteins, Animals, Humans, lcsh:Science, Polymerase, Multidisciplinary, 030102 biochemistry & molecular biology, biology, lcsh:R, Substrate (chemistry), [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, DNA, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], 030104 developmental biology, Monomer, chemistry, DNA repair enzymes, Enzyme mechanisms, Biophysics, biology.protein, Recombinant DNA, Cattle, lcsh:Q
Abstract

DNA-dependent poly(ADP-ribose) polymerases (PARPs) PARP1, PARP2 and PARP3 act as DNA break sensors signalling DNA damage. Upon detecting DNA damage, these PARPs use nicotine adenine dinucleotide as a substrate to synthesise a monomer or polymer of ADP-ribose (MAR or PAR, respectively) covalently attached to the acceptor residue of target proteins. Recently, it was demonstrated that PARP1–3 proteins can directly ADP-ribosylate DNA breaks by attaching MAR and PAR moieties to terminal phosphates. Nevertheless, little is still known about the mechanisms governing substrate recognition and specificity of PARP1, which accounts for most of cellular PARylation activity. Here, we characterised PARP1-mediated DNA PARylation of DNA duplexes containing various types of breaks at different positions. The 3′-terminal phosphate residue at double-strand DNA break ends served as a major acceptor site for PARP1-catalysed PARylation depending on the orientation and distance between DNA strand breaks in a single DNA molecule. A preference for ADP-ribosylation of DNA molecules containing 3′-terminal phosphate over PARP1 auto-ADP-ribosylation was observed, and a model of DNA modification by PARP1 was proposed. Similar results were obtained with purified recombinant PARP1 and HeLa cell-free extracts. Thus, the biological effects of PARP-mediated ADP-ribosylation may strongly depend on the configuration of complex DNA strand breaks.

Published
2020

9. FANCD2 modulates the mitochondrial stress response to prevent common fragile site instability

Author

Claude Saint-Ruf, Viviana Barra, Silvia Ravera, Philippe Fernandes, Viola Nähse, Enrico Cappelli, Valeria Naim, Benoit Miotto, Maha Said, Intégrité du génome et cancers (IGC), Institut Gustave Roussy (IGR)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Università degli studi di Palermo - University of Palermo, The Norwegian Radium Hospital [Oslo, Norway] (TNRH), University of Genoa (UNIGE), Istituto Giannina Gaslini, Genova, Immunologia Clinica e Sperimentale, École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Gustave Roussy (IGR)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Università degli studi di Genova = University of Genoa (UniGe), Miotto, Benoit, Fernandes P., Miotto B., Saint-Ruf C., Said M., Barra V., Nahse V., Ravera S., Cappelli E., and Naim V.

Subjects
0301 basic medicine, Genome instability, musculoskeletal diseases, Transcription, Genetic, QH301-705.5, Regulator, Medicine (miscellaneous), Mitochondrion, Biology, [SDV.BBM.BM] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, General Biochemistry, Genetics and Molecular Biology, Oxidative Phosphorylation, Article, 03 medical and health sciences, 0302 clinical medicine, Transcription (biology), Stress, Physiological, hemic and lymphatic diseases, Gene expression, FANCD2, Humans, Biology (General), Gene, Ubiquitins, Chromosomal fragile site, Chromosome Fragile Sites, Chromosome Fragility, Fanconi Anemia Complementation Group D2 Protein, DNA damage and repair, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, HCT116 Cells, Cell biology, Mitochondria, Settore BIO/18 - Genetica, 030104 developmental biology, Gene Expression Regulation, 030220 oncology & carcinogenesis, Unfolded Protein Response, General Agricultural and Biological Sciences, DNA Damage
Abstract

Common fragile sites (CFSs) are genomic regions frequently involved in cancer-associated rearrangements. Most CFSs lie within large genes, and their instability involves transcription- and replication-dependent mechanisms. Here, we uncover a role for the mitochondrial stress response pathway in the regulation of CFS stability in human cells. We show that FANCD2, a master regulator of CFS stability, dampens the activation of the mitochondrial stress response and prevents mitochondrial dysfunction. Genetic or pharmacological activation of mitochondrial stress signaling induces CFS gene expression and concomitant relocalization to CFSs of FANCD2. FANCD2 attenuates CFS gene transcription and promotes CFS gene stability. Mechanistically, we demonstrate that the mitochondrial stress-dependent induction of CFS genes is mediated by ubiquitin-like protein 5 (UBL5), and that a UBL5-FANCD2 dependent axis regulates the mitochondrial UPR in human cells. We propose that FANCD2 coordinates nuclear and mitochondrial activities to prevent genome instability., Fernandes et al. discover a connection between the mitochondrial stress response and genomic stability. They find that transcription of common fragile site (CFS) genes is induced by mitochondrial stress, whereas a regulator of CFS stability, FANCD2, acts to dampen the mitochondrial stress response and transcription-associated replication stress. These findings suggest a FANCD2-mediated coordination of nuclear and mitochondrial responses to stress.

Published
2021

10. Unscheduled origin building in S-phase upon tight CDK1 inhibition suppresses CFS instability

Author

Michelle Debatisse, El-Hilali S, Azar D, A.-M. Lachages, Yan Jaszczyszyn, Chun-Long Chen, Mélanie Schmidt, Stefano Gnan, Claude Thermes, Koundrioukoff S, Olivier Brison, Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Dynamique de l'information génétique : bases fondamentales et cancer (DIG CANCER), Centre National de la Recherche Scientifique (CNRS)-Institut Curie [Paris]-Sorbonne Université (SU), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Unité de Technologies Chimiques et Biologiques pour la Santé (UTCBS - UM 4 (UMR 8258 / U1022)), Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Institut Curie [Paris]-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Intégrité du génome et cancers (IGC), École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Gustave Roussy (IGR)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), École pratique des hautes études (EPHE), and CHEN, Chunlong

Subjects
[SDV.CAN]Life Sciences [q-bio]/Cancer, [SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], Biology, [SDV.GEN.GH] Life Sciences [q-bio]/Genetics/Human genetics, [SDV.BBM.BM] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Instability, Chromosome segregation, DNA replication factor CDT1, 03 medical and health sciences, 0302 clinical medicine, [SDV.CAN] Life Sciences [q-bio]/Cancer, Transcription (biology), [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], [SDV.BC.BC] Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], Mitosis, 030304 developmental biology, 0303 health sciences, Cyclin-dependent kinase 1, Chromosomal fragile site, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Cell biology, [SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics, 030220 oncology & carcinogenesis, biology.protein, [SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Chromosome breakage
Abstract

SummaryGenome integrity requires replication to be completed before chromosome segregation. This coordination essentially relies on replication-dependent activation of a dedicated checkpoint that inhibits CDK1, delaying mitotic onset. Under-replication of Common Fragile Sites (CFSs) however escapes surveillance, which triggers chromosome breakage. Using human cells, we asked here whether such leakage results from insufficient CDK1 inhibition under modest stresses used to destabilize CFSs. We found that tight CDK1 inhibition suppresses CFS instability. Repli-Seq and molecular combing analyses consistently showed a burst of replication initiations in mid S phase across large origin-poor domains shaped by transcription, including CFSs. Strikingly, CDC6 or CDT1 depletion or CDC7-DBF4 inhibition during the S phase prevented both extra-initiations and CFS rescue, showing that CDK1 inhibition promotes targeted and mistimed building of functional extra-origins. In addition to delay mitotic onset, checkpoint activation therefore advances replication completion of chromosome domains at risk of under-replication, two complementary roles preserving genome stability.

Published
2021

11. Fanconi anemia proteins counteract the implementation of the oncogene-induced senescence program

Author

Caroline Evrard, Françoise Dessarps-Freichey, Anne Helbling-Leclerc, Filippo Rosselli, Physiopathologie et thérapie du muscle strié, Université Pierre et Marie Curie - Paris 6 (UPMC)-IFR14-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de Recherche Jean-Pierre AUBERT Neurosciences et Cancer - U1172 Inserm - U837 (JPArc), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Lille Nord de France (COMUE)-Université de Lille, Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Centre de Recherche Jean-Pierre AUBERT Neurosciences et Cancer - U837 (JPArc), and Université Lille Nord de France (COMUE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille

Subjects
DNA Replication, Senescence, congenital, hereditary, and neonatal diseases and abnormalities, DNA Repair, Cathepsin L, [SDV]Life Sciences [q-bio], lcsh:Medicine, Article, Cell Line, 03 medical and health sciences, 0302 clinical medicine, Ubiquitin, Downregulation and upregulation, Fanconi anemia, hemic and lymphatic diseases, FANCD2, medicine, Humans, Monoubiquitination, lcsh:Science, Cellular Senescence, ComputingMilieux_MISCELLANEOUS, Cell Proliferation, 030304 developmental biology, 0303 health sciences, Multidisciplinary, Fanconi Anemia Complementation Group A Protein, biology, Oncogene, Fanconi Anemia Complementation Group D2 Protein, lcsh:R, Ubiquitination, nutritional and metabolic diseases, Oncogenes, medicine.disease, FANCA, Cell Transformation, Neoplastic, Fanconi Anemia, 030220 oncology & carcinogenesis, Cancer research, biology.protein, lcsh:Q, Reactive Oxygen Species, DNA Damage
Abstract

Fanconi Anemia (FA), due to the loss-of-function of the proteins that constitute the FANC pathway involved in DNA replication and genetic stability maintainance, is a rare genetic disease featuring bone marrow failure, developmental abnormalities and cancer predisposition. Similar clinical stigmas have also been associated with alterations in the senescence program, which is activated in physiological or stress situations, including the unscheduled, chronic, activation of an oncogene (oncogene induced senescence, OIS). Here, we wanted to determine the crosstalk, if any, between the FANC pathway and the OIS process. OIS was analyzed in two known cellular models, IMR90-hTERT/ER:RASG12V and WI38-hTERT/ER:GFP:RAF1, harboring 4-hydroxytamoxifen-inducible oncogenes. We observed that oncogene activation induces a transitory increase of both FANCA and FANCD2 as well as FANCD2 monoubiquitination, readout of FANC pathway activation, followed by their degradation. FANCD2 depletion, which leads to a pre-senescent phenotype, anticipates OIS progression. Coherently, FANCD2 overexpression or inhibition of its proteosomal-dependent degradation slightly delays OIS progression. The pro-senescence protease cathepsin L, which activation is anticipated during OIS in FANCD2-depleted cells, also participates to FANCD2 degradation. Our results demonstrate that oncogene activation is first associated with FANCD2 induction and activation, which may support initial cell proliferation, followed by its degradation/downregulation when OIS proceeds.

Published
2019

12. A Recurrent Activating Missense Mutation in Waldenström Macroglobulinemia Affects the DNA Binding of the ETS Transcription Factor SPI1 and Enhances Proliferation

Author

Nathalie Droin, Philippe Rameau, Philippe Dessen, Florence Nguyen-Khac, Bilyana Stoilova, Elena Mylonas, Marine Armand, Damien Roos-Weil, Olivier Bernard, Els Verhoeyen, Paresh Vyas, Said Aoufouchi, Marlen Metzner, Françoise Pflumio, Frédéric Charlotte, Vincent Ribrag, Diane Lara, Olivier Elemento, Pierre Morel, Eric Durot, Cécile Hérate, Pascale Cornillet-Lefebvre, Véronique Della-Valle, Hussein Ghamlouch, Virginie Ropars, Frederik Damm, Nabih Azar, Christel Guillouf, François-Loïc Cosset, Camille Decaudin, Valérie Camara-Clayette, M'Boyba Diop, Veronique Leblond, Thomas Mercher, Jean-Baptiste Charbonnier, Rima Haddad, Hématopoïèse normale et pathologique (U1170 Inserm), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Enveloppe Nucléaire, Télomères et Réparation de l’ADN (INTGEN), Département Biochimie, Biophysique et Biologie Structurale (B3S), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université Paris Descartes - Faculté de Médecine (UPD5 Médecine), Université Paris Descartes - Paris 5 (UPD5), Department of Haematology, Haemostasis, Oncology and Stem Cell Transplantation (MHH), Hannover Medical School [Hannover] (MHH), CEA Direction des Sciences du Vivant (CEA/DSV), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), The Weatherall Institute of Molecular Medicine, University of Oxford [Oxford], Weill Medical College of Cornell University [New York], Institut Gustave Roussy (IGR), Plateforme de Bioinformatique [Gustave Roussy], Analyse moléculaire, modélisation et imagerie de la maladie cancéreuse (AMMICa), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de recherche translationnelle (Labo RT), Immunologie des tumeurs et immunothérapie (UMR 1015), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Virus enveloppés, vecteurs et immunothérapie – Enveloped viruses, Vectors and Immuno-therapy (EVIR), Centre International de Recherche en Infectiologie - UMR (CIRI), École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM), CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Service d'Hématologie clinique [CHU Pitié-Salpêtrière], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Sorbonne Université (SU), Département d'hématologie [Gustave Roussy], Laboratoire d'hématologie, Centre Hospitalier Universitaire de Reims (CHU Reims), Plateforme imagerie et cytométrie (PFIC), Ligue Nationale Contre le Cancer - Paris, Ligue Nationale Contre le Cancer (LNCC), Brigham and Women's Hospital [Boston], Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Service d'Hématologie Biologique [CHU Pitié-Salpêtrière], Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Gustave Roussy (IGR)-Université Paris-Sud - Paris 11 (UP11)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Gustave Roussy (IGR)-Université Paris-Sud - Paris 11 (UP11), Institut National de la Santé et de la Recherche Médicale (INSERM)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Ligue Nationnale Contre le Cancer, Centre de Recherche et d'Etude en Droit et Science Politique (CREDESPO), Université de Bourgogne (UB), University of Oxford, Centre International de Recherche en Infectiologie (CIRI), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and École Pratique des Hautes Études (EPHE)

Subjects
0301 basic medicine, [SDV]Life Sciences [q-bio], Mutation, Missense, lymphoma, Biology, Cell Line, Mice, 03 medical and health sciences, 0302 clinical medicine, Germline mutation, Mutant protein, Proto-Oncogene Proteins, spi-1/pu.1, pu.1, Animals, Humans, somatic mutation, Nucleotide Motifs, Lenalidomide, Transcription factor, mouse, Cell Proliferation, cll, B-Lymphocytes, Binding Sites, SPI1, Base Sequence, Proto-Oncogene Proteins c-ets, Point mutation, ETS transcription factor family, Promoter, Azepines, Triazoles, Cell biology, cell differentiation, 030104 developmental biology, Gene Expression Regulation, Oncology, 030220 oncology & carcinogenesis, Myeloid Differentiation Factor 88, Trans-Activators, acute myeloid-leukemia, Waldenstrom Macroglobulinemia, Transcription Factor Gene, Protein Binding, Transcription Factors
Abstract

The ETS-domain transcription factors divide into subfamilies based on protein similarities, DNA-binding sequences, and interaction with cofactors. They are regulated by extracellular clues and contribute to cellular processes, including proliferation and transformation. ETS genes are targeted through genomic rearrangements in oncogenesis. The PU.1/SPI1 gene is inactivated by point mutations in human myeloid malignancies. We identified a recurrent somatic mutation (Q226E) in PU.1/SPI1 in Waldenström macroglobulinemia, a B-cell lymphoproliferative disorder. It affects the DNA-binding affinity of the protein and allows the mutant protein to more frequently bind and activate promoter regions with respect to wild-type protein. Mutant SPI1 binding at promoters activates gene sets typically promoted by other ETS factors, resulting in enhanced proliferation and decreased terminal B-cell differentiation in model cell lines and primary samples. In summary, we describe oncogenic subversion of transcription factor function through subtle alteration of DNA binding leading to cellular proliferation and differentiation arrest. Significance: The demonstration that a somatic point mutation tips the balance of genome-binding pattern provides a mechanistic paradigm for how missense mutations in transcription factor genes may be oncogenic in human tumors. This article is highlighted in the In This Issue feature, p. 681

Published
2019

13. Transcription-dependent regulation of replication dynamics modulates genome stability

Author

Marion Blin, Caroline Brossas, Gaël A. Millot, Mélanie Schmidt, Viola Nähse, Marie-Noëlle Prioleau, Michelle Debatisse, Benoît Le Tallec, Dynamique de l'information génétique : bases fondamentales et cancer (DIG CANCER), Centre National de la Recherche Scientifique (CNRS)-Institut Curie [Paris]-Sorbonne Université (SU), Centre de Recherche en Cancérologie de Marseille (CRCM), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Aix Marseille Université (AMU), Institute for Cancer Research [Oslo], Oslo University Hospital [Oslo], Institute of Clinical Medicine [Oslo], Faculty of Medicine [Oslo], University of Oslo (UiO)-University of Oslo (UiO), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Institut Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Hub Bioinformatique et Biostatistique - Bioinformatics and Biostatistics HUB, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), The M.D. team is supported by the Agence Nationale de la Recherche (grant ANR-13-BSV6-0008-01/FRA-Dom), the Association pour la Recherche sur le Cancer (grant Subvention Libre Sl220130607073), and the Institut National du Cancer (grants INCa subventions 2013-103 and PLBIO17-194). The M.N.P. team is supported by the Association pour la Recherche sur le Cancer (grant Labellisation PGA120150202272) and the Agence Nationale de la Recherche (grant ANR-15-CE12-0004-01). M.B. was supported by fellowships from the Ministère de l’Enseignement Supérieur et de la Recherche and the Ligue contre le cancer., We thank S. Lambert and O. Hyrien for critical reading of the manuscript. The authors would like to acknowledge the Cell and Tissue Imaging Platform – PICT-IBiSA (member of France-Bioimaging) of the Genetics and Developmental Biology Department (UMR3215/U934) of Institut Curie for help with light microscopy, the Flow Cytometry Platform Imagoseine of Institut Jacques Monod, Université Paris Diderot, and the Imaging and Cytometry Platform (PFIC) of Institut Gustave Roussy for assistance with cell sorting., ANR-13-BSV6-0008,FRA-Dom,Chorégraphie des domaines de la chromatine au cours de la différenciation: réorganisation des profils de réplication et des sites fragiles communs(2013), Institut Curie [Paris]-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Aix Marseille Université (AMU)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut de biologie de l'ENS Paris (UMR 8197/1024) (IBENS), Département de Biologie - ENS Paris, École normale supérieure - Paris (ENS Paris)-École normale supérieure - Paris (ENS Paris)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut Curie-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Université de Lorraine (UL), Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7), Université Paris Descartes - Faculté de Chirurgie Dentaire (UPD5 Odontologie), Université Paris Descartes - Paris 5 (UPD5), and Institut Curie

Subjects
DNA Replication, MESH: Genomic Instability / physiology, Transcription, Genetic, [SDV]Life Sciences [q-bio], Mitosis, [SDV.CAN]Life Sciences [q-bio]/Cancer, Endogeny, Biology, Genomic Instability, 03 medical and health sciences, 0302 clinical medicine, Structural Biology, Transcription (biology), MESH: Transcription, Genetic / genetics, MESH: DNA Replication / genetics, Animals, Humans, MESH: Animals, MESH: Chromosome Fragile Sites / genetics, MESH: Genomic Instability / genetics, Molecular Biology, Gene, 030304 developmental biology, Genome stability, 0303 health sciences, Replication timing, MESH: Humans, MESH: Mitosis / physiology, Chromosome Fragile Sites, Chromosomal fragile site, Promoter, Cell biology, MESH: Mitosis / genetics, MESH: Chromosome Fragile Sites / physiology, Fragile sites, MESH: DNA Replication / physiology, Transcription, Origin selection, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology, 030217 neurology & neurosurgery
Abstract

International audience; Common fragile sites (CFSs) are loci that are hypersensitive to replication stress and hotspots for chromosomal rearrangements in cancers. CFSs replicate late in S phase, are cell-type specific and nest in large genes. The relative impact of transcription-replication conflicts versus a low density in initiation events on fragility is currently debated. Here we addressed the relationships between transcription, replication, and instability by manipulating the transcription of endogenous large genes in chicken and human cells. We found that inducing low transcription with a weak promoter destabilized large genes, whereas stimulating their transcription with strong promoters alleviated instability. Notably, strong promoters triggered a switch to an earlier replication timing, supporting a model in which high transcription levels give cells more time to complete replication before mitosis. Transcription could therefore contribute to maintaining genome integrity, challenging the dominant view that it is exclusively a threat.

Published
2018

14. Epigenomic signature of the progeroid Cockayne syndrome exposes distinct and common features with physiological ageing

Author

Steve Horvath, Maria Giulia Bacalini, Claudio Franceschi, Paolo Garagnani, Claudia Chica, Miria Ricchetti, Clément Crochemore, Giovanna Lattanzi, Alain Sarasin, Cellules Souches et Développement / Stem Cells and Development, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Sup'Biotech, Hub Bioinformatique et Biostatistique - Bioinformatics and Biostatistics HUB, Karolinska Institutet [Stockholm], University hospital - Policlinico S.Orsola-Malpighi [Bologna, Italy], CNR Institute of Molecular Genetics 'Luigi Luca Cavalli-Sforza', Istituto Ortopedico Rizzoli [Bologna, Italy], University of California (UC), Institut Gustave Roussy (IGR), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Lobachevsky State University [Nizhni Novgorod], IRCCS Istituto delle Scienze Neurologiche di Bologna [Bologna, Italy], Ospedale Bellaria [Bologna, Italy], This work was supported by Agence Nationale de la Recherche (grant CS_AGE, aapg2019), DARRI (Institut Pasteur R&D, grant DISAGE, PasteurInnov2014), Programmes Transversales de Recherche, Institut Pasteur (grant PTR111-2017), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), and University of California

Subjects
Genetics, 0303 health sciences, Mutation, DNA repair, [SDV]Life Sciences [q-bio], dNaM, Biology, medicine.disease, medicine.disease_cause, Phenotype, Cockayne syndrome, 3. Good health, 03 medical and health sciences, 0302 clinical medicine, DNA methylation, medicine, Epigenetics, 030217 neurology & neurosurgery, 030304 developmental biology, Epigenomics
Abstract

Cockayne syndrome (CS) and UV-sensitivity syndrome (UVSS) are rare genetic disorders caused by mutation of the DNA repair and chromatin remodelling proteins CSA or CSB, but only CS patients display a progeroid and neurodegenerative phenotype. As epigenetic modifications constitute a well-established hallmark of ageing, we characterized genome-wide DNA methylation (DNAm) of fibroblasts from CS versus UVSS patients and healthy donors. The analysis of differentially methylated positions and regions revealed a CS-specific epigenetic signature, enriched in developmental transcription factors, transmembrane transporters, and cell adhesion factors. The CS-specific signature compared to DNAm changes in other progeroid diseases and regular ageing, identifyied commonalities and differences in epigenetic remodelling. CS shares DNAm changes with normal ageing more than other progeroid diseases do, and according to the methylation clock CS samples show up to 13-fold accelerated ageing. Thus, CS is characterized by a specific epigenomic signature that partially overlaps with and exacerbates DNAm changes occurring in physiological aging. Our results unveil new genes and pathways that are potentially relevant for the progeroid/degenerative CS phenotype.

Published
2021

15. Association of AXL and PD-L1 Expression with Clinical Outcomes in Patients with Advanced Renal Cell Carcinoma Treated with PD-1 Blockade

Author

Gro Gausdal, Frédéric Dugay, Salem Chouaib, Wolf H. Fridman, Yann Vano, Maxime Meylan, Julien Adam, Stéphane Terry, Alexandra Lespagnol, James B. Lorens, Sylvie Chabaud, Fathia Mami-Chouaib, C. Dalban, Benoit Beuselinck, Stéphanie Buart, Nathalie Rioux-Leclercq, Bernard Escudier, Laurence Albiges, Nathalie Chaput, Janice Barros-Monteiro, Guillaume Lacroix, Irelka Colina Moreno, Catherine Sautès-Fridman, Antoine Bougoüin, Immunologie intégrative des tumeurs et immunothérapie des cancers (INTIM), Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay, Centre Léon Bérard [Lyon], Institut de recherche en santé, environnement et travail (Irset), Université d'Angers (UA)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), CHU Pontchaillou [Rennes], Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université de Paris (UP), École des Hautes Études en Santé Publique [EHESP] (EHESP), University of Bergen (UiB), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Analyse moléculaire, modélisation et imagerie de la maladie cancéreuse (AMMICa), Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Catholic University of Leuven - Katholieke Universiteit Leuven (KU Leuven), Jonchère, Laurent, Université d'Angers (UA)-Université de Rennes (UR)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Cité (UPCité), NSERM, Université Paris-Saclay, Université de Paris, EL2015.LNCC/SaC, la Ligue Contre le Cancer, INCa-DGOS-PRT-K16–181, l'Institut National du Cancer and Direction générale de l'offre de soins, CARPEM program of the Sites Intégrés de Recherche sur le Cancer, Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de la Santé et de la Recherche Médicale (INSERM)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Université d'Angers (UA), and École Pratique des Hautes Études (EPHE)

Subjects
PD-L1, Cancer Research, [SDV]Life Sciences [q-bio], Programmed Cell Death 1 Receptor, Receptor tyrosine kinase, B7-H1 Antigen, 03 medical and health sciences, 0302 clinical medicine, Renal cell carcinoma, VHL, anti-PD- 1, medicine, Humans, Lung cancer, Carcinoma, Renal Cell, 030304 developmental biology, CD20, 0303 health sciences, biology, business.industry, AXL, Cancer, medicine.disease, Kidney Neoplasms, clear-cell renal cell carcinoma, 3. Good health, Blockade, [SDV] Life Sciences [q-bio], Vascular endothelial growth factor A, Nivolumab, Oncology, 030220 oncology & carcinogenesis, biology.protein, Cancer research, immunotherapy, business
Abstract

Purpose: A minority of patients currently respond to single-agent immune-checkpoint blockade (ICB), and strategies to increase response rates are urgently needed. AXL is a receptor tyrosine kinase commonly associated with drug resistance and poor prognosis in many cancer types, including in clear-cell renal cell carcinoma (ccRCC). Recent experimental cues in breast, pancreatic, and lung cancer models have linked AXL with immune suppression and resistance to antitumor immunity. However, its role in intrinsic and acquired resistance to ICB remains largely unexplored. Experimental Design: In this study, tumoral expression of AXL was examined in ccRCC specimens from 316 patients who were metastatic receiving the PD-1 inhibitor nivolumab in the GETUG AFU 26 NIVOREN trial after failure of antiangiogenic therapy. We assessed associations between AXL and patient outcomes following PD-1 blockade, as well as the relationship with various markers, including PD-L1; VEGFA; the immune markers CD3, CD8, CD163, and CD20; and the mutational status of the tumor-suppressor gene von Hippel-Lindau (VHL). Results: Our results show that high AXL-expression level in tumor cells is associated with lower response rates and a trend to shorter progression-free survival following anti–PD-1 treatment. AXL expression was strongly associated with tumor–PD-L1 expression, especially in tumors with VHL inactivation. Moreover, patients with tumors displaying concomitant PD-L1 expression and high AXL expression had the worst overall survival. Conclusions: Our findings propose AXL as candidate factor of resistance to PD-1 blockade, and provide compelling support for screening both AXL and PD-L1 expression in the management of advanced ccRCC. See related commentary by Hahn et al., p. 6619

Published
2021

16. The Telomeric Protein TRF2 Regulates Replication Origin Activity within Pericentromeric Heterochromatin

Author

Jing Ye, Serge Bauwens, Liudmyla Lototska, Stephane Koundrioukoff, Michelle Debatisse, Aaron Mendez-Bermudez, Eric Gilson, Institut de Recherche sur le Cancer et le Vieillissement (IRCAN), Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Côte d'Azur (UCA), Institut Gustave Roussy (IGR), Intégrité du génome et cancers (IGC), Institut Gustave Roussy (IGR)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), and Shanghai Jiao Tong University [Shanghai]

Subjects
replication, Heterochromatin, Satellite DNA, pericentromeric DNA, TRF2, Origin of replication, General Biochemistry, Genetics and Molecular Biology, Article, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], lcsh:Science, ORC2, Ecology, Evolution, Behavior and Systematics, 030304 developmental biology, 0303 health sciences, biology, heterochromatin, Paleontology, Helicase, telomeres, Chromatin, Cell biology, chemistry, ORC, Space and Planetary Science, Replication Initiation, biology.protein, lcsh:Q, 030217 neurology & neurosurgery, DNA
Abstract

International audience; Heterochromatic regions render the replication process particularly difficult due to the high level of chromatin compaction and the presence of repeated DNA sequences. In humans, replication through pericentromeric heterochromatin requires the binding of a complex formed by the telomeric factor TRF2 and the helicase RTEL1 in order to relieve topological barriers blocking fork progression. Since TRF2 is known to bind the Origin Replication Complex (ORC), we hypothesized that this factor could also play a role at the replication origins (ORI) of these heterochromatin regions. By performing DNA combing analysis, we found that the ORI density is higher within pericentromeric satellite DNA repeats than within bulk genomic DNA and decreased upon TRF2 downregulation. Moreover, we showed that TRF2 and ORC2 interact in pericentromeric DNA, providing a mechanism by which TRF2 is involved in ORI activity. Altogether, our findings reveal an essential role for TRF2 in pericentromeric heterochromatin replication by regulating both replication initiation and elongation.

Published
2021

17. Role of the BAHD1 Chromatin-Repressive Complex in Placental Development and Regulation of Steroid Metabolism

Author

Slimane Ait-Si-Ali, Marie Wattenhofer-Donzé, Marie-France Champy, Renaud Pourpre, Alice Lebreton, Goran Lakisic, Pascale Cossart, Morwenna Le Guillou, Hélène Bierne, Tania Sorg, Guillaume Soubigou, Emanuele Libertini, Anne C. Ferguson-Smith, Jean Feunteun, Jean-Yves Coppée, Elizabeth J. Radford, Olivia Wendling, MICrobiologie de l'ALImentation au Service de la Santé (MICALIS), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, Interactions Bactéries-Cellules (UIBC), Institut National de la Recherche Agronomique (INRA)-Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Clinique de la Souris (ICS), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Transcriptome et Epigénome (PF2), Institut Pasteur [Paris] (IP), Cambridge University Hospitals - NHS (CUH), University of Cambridge [UK] (CAM), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Centre épigénétique et destin cellulaire (EDC), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Department of Genetics, French Ligue Nationale Contre le Cancer (comité régional d’Ile–de-France, LNCC RS10/75–76 and LNCC 131/12, INRA AO blanc MICA 2011, iXcore Fundation for Research, ANR-11-BSV3-0003,EPILIS,Reprogrammation épigénétique par la bactérie pathogène Listeria monocytogenes(2011), European Project: 670823,H2020,ERC-2014-ADG,BacCellEpi(2015), Institut National de la Recherche Agronomique (INRA)-Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pasteur [Paris], Centre épigénétique et destin cellulaire (EDC (UMR_7216)), Lebreton, Alice, BLANC - Reprogrammation épigénétique par la bactérie pathogène Listeria monocytogenes - - EPILIS2011 - ANR-11-BSV3-0003 - BLANC - VALID, Bacterial, cellular and epigenetic factors that control enteropathogenicity - BacCellEpi - - H20202015-10-01 - 2018-09-30 - 670823 - VALID, Bierne, Hélène, Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur [Paris]-Institut National de la Recherche Agronomique (INRA), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Radford, Elizabeth [0000-0002-8336-6045], Ferguson-Smith, Anne [0000-0003-4996-9990], and Apollo - University of Cambridge Repository

Subjects
0301 basic medicine, Embryology, Cancer Research, Chromosomal Proteins, Non-Histone, Placenta, dna methylation, [SDV.GEN] Life Sciences [q-bio]/Genetics, [SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], [SDV.BBM.BM] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Biochemistry, Mice, Pregnancy, Gene expression, Medicine and Health Sciences, Small interfering RNAs, Genetics (clinical), estrogen-receptor-alpha, histone deacetylase, transcriptional repression, glycogen cells, breast-cancer, sant domain, Regulation of gene expression, Mice, Knockout, biology, Chromosome Biology, [SDV.BDD.EO] Life Sciences [q-bio]/Development Biology/Embryology and Organogenesis, Gene Expression Regulation, Developmental, Nuclear Proteins, Genomics, Chromatin, Precipitation Techniques, Cell biology, Nucleic acids, DNA-Binding Proteins, Histone, DNA methylation, [SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Epigenetics, Female, Steroids, Anatomy, DNA modification, Transcriptome Analysis, Chromatin modification, Research Article, lcsh:QH426-470, Steroid hormone receptor, Research and Analysis Methods, 03 medical and health sciences, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], [SDV.BC.BC] Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], Genetics, Immunoprecipitation, Animals, Humans, Non-coding RNA, Molecular Biology, Transcription factor, Ecology, Evolution, Behavior and Systematics, [SDV.GEN]Life Sciences [q-bio]/Genetics, Reproductive System, Estrogen Receptor alpha, Biology and Life Sciences, Computational Biology, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Cell Biology, DNA, Genome Analysis, Molecular biology, Placentation, Gene regulation, lcsh:Genetics, 030104 developmental biology, [SDV.BDD.EO]Life Sciences [q-bio]/Development Biology/Embryology and Organogenesis, HEK293 Cells, biology.protein, RNA, Transcriptome, Developmental Biology, Transcription Factors
Abstract

BAHD1 is a vertebrate protein that promotes heterochromatin formation and gene repression in association with several epigenetic regulators. However, its physiological roles remain unknown. Here, we demonstrate that ablation of the Bahd1 gene results in hypocholesterolemia, hypoglycemia and decreased body fat in mice. It also causes placental growth restriction with a drop of trophoblast glycogen cells, a reduction of fetal weight and a high neonatal mortality rate. By intersecting transcriptome data from murine Bahd1 knockout (KO) placentas at stages E16.5 and E18.5 of gestation, Bahd1-KO embryonic fibroblasts, and human cells stably expressing BAHD1, we also show that changes in BAHD1 levels alter expression of steroid/lipid metabolism genes. Biochemical analysis of the BAHD1-associated multiprotein complex identifies MIER proteins as novel partners of BAHD1 and suggests that BAHD1-MIER interaction forms a hub for histone deacetylases and methyltransferases, chromatin readers and transcription factors. We further show that overexpression of BAHD1 leads to an increase of MIER1 enrichment on the inactive X chromosome (Xi). In addition, BAHD1 and MIER1/3 repress expression of the steroid hormone receptor genes ESR1 and PGR, both playing important roles in placental development and energy metabolism. Moreover, modulation of BAHD1 expression in HEK293 cells triggers epigenetic changes at the ESR1 locus. Together, these results identify BAHD1 as a core component of a chromatin-repressive complex regulating placental morphogenesis and body fat storage and suggest that its dysfunction may contribute to several human diseases., Author Summary The importance of epigenetics in regulation and dysfunction of metabolic pathways is increasingly recognized but the underlying mechanisms and molecular actors involved remain incompletely characterized. Here, we provide evidence that the heterochromatinization factor BAHD1 cooperates with MIER proteins to assemble chromatin-repressive complexes that control a network of metabolic genes involved in placental and fetal growth and in cholesterol homeostasis.

Published
2020
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18. Les poly(ADP-Ribose) polymérases 1 et 2 d'Arabidopsis thaliana modifient l'ADN en libérant des résidus terminaux de phosphate

Author

Taipakova, Sabira, Kuanbay, Aigerim, Saint-Pierre, Christine, Gasparutto, Didier, Baiken, Yeldar, Groisman, Regina, Ishchenko, Alexander, Saparbaev, Murat, Bissenbaev, Amangeldy, Laboratoire Epigenetique et Cancer, Centre National de la Recherche Scientifique (CNRS), Réparation de l’ADN (CNRS UMR 8200), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Institut Gustave Roussy (IGR), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), and Al-Farabi Kazakh National University

Subjects
target of rapamycin, Torin1, rapamycin, [SDV]Life Sciences [q-bio], food and beverages, alpha-amylase, TOR, Wheat seed, Abscisic acid, TOR pathway, [CHIM]Chemical Sciences, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Gibberellic acid, S6 kinase 1
Abstract

International audience; Germination is a process of seed sprouting that facilitates embryo growth. The breakdown of reserved starch in the endosperm into simple sugars is essential for seed germination and subsequent seedling growth. At the early stage of germination, gibberellic acid (GA) activates transcription factor GAMYB to promote de novo synthesis of isoforms of α‐amylase in the aleurone layer and scutellar epithelium of the embryo. Here, we demonstrate that wheat germination is regulated by plant target of rapamycin signaling. TOR is a central component of the essential-nutrient–dependent pathway controlling cell growth in all eukaryotes. It is known that rapamycin, a highly specific allosteric inhibitor of TOR, is effective in yeast and animal cells but ineffective in most of higher plants likely owing to structural differences in ubiquitous rapamycin receptor FKBP12. The action of rapamycin on wheat growth has not been studied. Our data show that rapamycin inhibits germination of wheat seeds and of their isolated embryos in a dose-dependent manner. The involvement of Triticum aestivum TOR (TaTOR) in wheat germination was consistent with the suppression of wheat embryo growth by specific inhibitors of the TOR kinase: pp242 or torin1. Rapamycin or torin1 interfered with GA function in germination because of a potent inhibitory effect on α-amylase and GAMYB gene expression. The TOR inhibitors selectively targeted the GA-dependent gene expression, whereas expression of the abscisic acid-dependent ABI5 gene was not affected by either rapamycin or torin1. To determine whether the TaTOR kinase activation takes place during wheat germination, we examined phosphorylation of a ribosomal protein, T. aestivum S6 kinase 1 (TaS6K1; a substrate of TOR). The phosphorylation of serine 467 (S467) in a hydrophobic motif on TaS6K1 was induced in a process of germination triggered by GA. Moreover, the germination-induced phosphorylation of TaS6K1 on S467 was dependent on TaTOR and was inhibited by rapamycin or torin1. Besides, a gibberellin biosynthesis inhibitor (paclobutrazol; PBZ) blocked not only α-amylase gene expression but also TaS6K1 phosphorylation in wheat embryos. Thus, a hormonal action of GA turns on the synthesis of α-amylase in wheat germination via activation of the TaTOR–S6K1 signaling pathway.; La germination est un processus de germination des graines qui facilite la croissance de l'embryon. La décomposition de l'amidon réservé dans l'endosperme en sucres simples est essentielle à la germination des graines et à la croissance ultérieure des semis. Au stade précoce de la germination, l'acide gibbérellique (AG) active le facteur de transcription GAMYB pour favoriser la synthèse de novo des isoformes de α-amylase dans la couche d'aleurone et l'épithélium scutellaire de l'embryon. Ici, nous démontrons que la germination du blé est régulée par la cible végétale de la signalisation de la rapamycine. Le TOR est un élément central de la voie dépendant des nutriments essentiels qui contrôle la croissance cellulaire chez tous les eucaryotes. On sait que la rapamycine, un inhibiteur allostérique hautement spécifique de TOR, est efficace dans les cellules de levure et les cellules animales, mais inefficace dans la plupart des plantes supérieures, probablement en raison de différences structurelles dans le récepteur omniprésent de la rapamycine FKBP12. L'action de la rapamycine sur la croissance du blé n'a pas été étudiée. Nos données montrent que la rapamycine inhibe la germination des graines de blé et de leurs embryons isolés de manière dose-dépendante. L'implication de la TOR de Triticum aestivum (TaTOR) dans la germination du blé était cohérente avec la suppression de la croissance des embryons de blé par des inhibiteurs spécifiques de la kinase TOR : pp242 ou torin1. La rapamycine ou torin1 a entravé la fonction de l'AG dans la germination en raison d'un puissant effet inhibiteur sur l'expression des gènes α-amylase et GAMYB. Les inhibiteurs de TOR ont ciblé sélectivement l'expression du gène GA-dépendant, alors que l'expression du gène ABI5 dépendant de l'acide abscissique n'a été affectée ni par la rapamycine ni par la torine1. Pour déterminer si l'activation de la kinase TaTOR a lieu pendant la germination du blé, nous avons examiné la phosphorylation d'une protéine ribosomique, la kinase 1 de T. aestivum S6 (TaS6K1 ; un substrat de TOR). La phosphorylation de la sérine 467 (S467) dans un motif hydrophobe sur TaS6K1 a été induite dans un processus de germination déclenché par l'AG. De plus, la phosphorylation de TaS6K1 induite par la germination sur S467 était dépendante de TaTOR et était inhibée par la rapamycine ou la torine1. En outre, un inhibiteur de la biosynthèse de la gibbérelline (paclobutrazol ; PBZ) a bloqué non seulement l'expression du gène α-amylase mais aussi la phosphorylation de TaS6K1 chez les embryons de blé. Ainsi, une action hormonale de l'AG active la synthèse de la α-amylase dans la germination du blé par l'activation de la voie de signalisation TaTOR-S6K1.Traduit avec www.DeepL.com/Translator (version gratuite)

Published
2020

19. La détection des évènements rares

Author

Idziorek, Thierry, Jouy, Nathalie, Touil, Yasmine, Ostyn, Pauline, Bio Imaging Center Lille, Université de Lille, Centre de Recherche Jean-Pierre AUBERT Neurosciences et Cancer - U1172 Inserm - U837 (JPArc), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Lille Nord de France (COMUE)-Université de Lille, Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Intégrité du génome et cancers (IGC), Institut Gustave Roussy (IGR)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Idziorek, Thierry, Plateforme BioImaging Center Lille (BICeL), Plateformes Lilloises en Biologie et Santé - UAR 2014 - US 41 (PLBS), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre National de la Recherche Scientifique (CNRS), Centre de Recherche Jean-Pierre AUBERT Neurosciences et Cancer - U837 (JPArc), Université Lille Nord de France (COMUE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille, École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Gustave Roussy (IGR)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), and École pratique des hautes études (EPHE)

Subjects
[SDV.BC]Life Sciences [q-bio]/Cellular Biology, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, ComputingMilieux_MISCELLANEOUS
Abstract

International audience

Published
2020

20. An Assay for the Activity of Base Excision Repair Enzymes in Cellular Extracts Using Fluorescent DNA Probes: ACTIVITY OF BASE EXCISION REPAIR ENZYMES

Author

Olga A. Kladova, Nikita A. Kuznetsov, Regina Groisman, Alexander A. Ishchenko, Danila A. Iakovlev, Murat Saparbaev, Olga S. Fedorova, Institute of Chemical Biology and Fundamental Medicine [Novosibirsk, Russia] (ICBFM SB RAS), Siberian Branch of the Russian Academy of Sciences (SB RAS), Intégrité du génome et cancers (IGC), Institut Gustave Roussy (IGR)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Novosibirsk State University (NSU), ANR-18-CE44-0008,DNAPAR18,Etude de l'ADP-ribosylation d'ADN et de son rôle dans la réponse aux dommages à l'ADN(2018), Réparation de l’ADN (CNRS UMR 8200), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), and Institut Gustave Roussy (IGR)

Subjects
Cell Extracts, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, DNA Repair, [SDV]Life Sciences [q-bio], AP-endonuclease, UDG, Biochemistry, AP endonuclease, DNA Glycosylases, Endonuclease, chemistry.chemical_compound, thymine DNA glycosylase, methyl-CpG-binding domain 4, AAG, OGG1, ComputingMilieux_MISCELLANEOUS, NTHL1, Cells, Cultured, 0303 health sciences, biology, Chemistry, DNA glycosylase, 030302 biochemistry & molecular biology, human AP endonuclease 1, General Medicine, Base excision repair, 3. Good health, NEIL1, MBD4, fluorescence, DNA Probes, human endonuclease III, 8-oxoguanosine, DNA damage, DNA repair, 6-carboxyfluorescein, BHQ, uracil-DNA glycosylase enzymatic activity, 8-oxoguanine DNA glycosylase, human endonuclease VIII, TDG, DHU, 03 medical and health sciences, Humans, AP site, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], alkyladenine DNA glycosylase, Enzyme Assays, Fluorescent Dyes, N6-ethenoadenosine, black hole quencher, 6-dihydrouridine, 3S)-2-(hydroxymethyl)-3-hydroxytetrahydrofuran residue, oxoG, FAM, εA, DNA probe, DNA Repair Enzymes, APE1, Förster resonance energy transfer, biology.protein, FRET, (2R, DNA, apurinic/apyrimidinic site, DNA Damage
Abstract

International audience; Damaged DNA bases are removed by the base excision repair (BER) mechanism. This enzymatic process begins with the action of one of DNA glycosylases, which recognize damaged DNA bases and remove them by hydrolyzing N-glycosidic bonds with the formation of apurinic/apyrimidinic (AP) sites. Apurinic/apyrimidinic endonuclease 1 (APE1) hydrolyzes the phosphodiester bond on the 5′-side of the AP site with generation of the single-strand DNA break. A decrease in the functional activity of BER enzymes is associated with the increased risk of cardiovascular, neurodegenerative, and oncological diseases. In this work, we developed a fluorescence method for measuring the activity of key human DNA glycosylases and AP endonuclease in cell extracts. The efficacy of fluorescent DNA probes was tested using purified enzymes; the most efficient probes were tested in the enzymatic activity assays in the extracts of A549, MCF7, HeLa, WT-7, HEK293T, and HKC8 cells. The activity of enzymes responsible for the repair of AP sites and removal of uracil and 5,6-dihydrouracil residues was higher in cancer cell lines as compared to the normal HKC8 human kidney cell line.; Les bases d'ADN endommagées sont éliminées par le mécanisme de réparation par excision de base (BER). Ce processus enzymatique commence par l'action d'une des glycosylases de l'ADN, qui reconnaît les bases d'ADN endommagées et les élimine en hydrolysant les liaisons N-glycosidiques avec la formation de sites apuriniques/apyrimidiniques (AP). L'endonucléase 1 apurinique /apyrimidinique (APE1) hydrolyse la liaison phosphodiester du côté 5′ du site AP avec la génération de la rupture de l'ADN simple brin. Une diminution de l'activité fonctionnelle des enzymes BER est associée à un risque accru de maladies cardiovasculaires, neurodégénératives et oncologiques. Dans ce travail, nous avons développé une méthode de fluorescence pour mesurer l'activité des principales glycosylases de l'ADN humain et de l'endonucléase AP dans les extraits cellulaires. L'efficacité des sondes d'ADN fluorescentes a été testée en utilisant des enzymes purifiées ; les sondes les plus efficaces ont été testées dans les essais d'activité enzymatique dans les extraits de cellules A549, MCF7, HeLa, WT-7, HEK293T, et HKC8. L'activité des enzymes responsables de la réparation des sites AP et de l'élimination des résidus d'uracile et de 5,6-dihydrouracile était plus élevée dans les lignées de cellules cancéreuses que dans la lignée normale de cellules rénales humaines HKC8.Traduit avec www.DeepL.com/Translator (version gratuite)

Published
2020

21. Biotechnological applications of the sepiolite interactions with bacteria: Bacterial transformation and DNA extraction

Author

Fidel Antonio Castro-Smirnov, Eduardo Ruiz-Hitzky, Olivier Piétrement, Pilar Aranda, Bernard S. Lopez, Eric Le Cam, Intégrité du génome et cancers (IGC), Institut Gustave Roussy (IGR)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Laboratoire Interdisciplinaire Carnot de Bourgogne [Dijon] (LICB), Université de Bourgogne (UB)-Université de Technologie de Belfort-Montbeliard (UTBM)-Centre National de la Recherche Scientifique (CNRS), Signalisation, noyaux et innovations en cancérologie (UMR8126), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Instituto de Ciencia de Materiales de Madrid (ICMM), Consejo Superior de Investigaciones Científicas [Madrid] (CSIC), Institut Gustave Roussy (IGR), [Institut Cochin] Département Développement, Reproduction et Cancer (DRC), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Gustave Roussy (IGR)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Laboratoire Interdisciplinaire Carnot de Bourgogne (ICB), Université de Technologie de Belfort-Montbeliard (UTBM)-Université de Bourgogne (UB)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Centre National de la Recherche Scientifique (CNRS), Aspects métaboliques et systémiques de l'oncogénèse pour de nouvelles approches thérapeutiques (METSY), Interactions moléculaires et cancer (IMC (UMR 8126)), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), PIETREMENT, Olivier, Stabilité Génétique et Oncogenèse (UMR 8200), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Laboratoire Interdisciplinaire Carnot de Bourgogne [Dijon] (ICB), and Centre National de la Recherche Scientifique (CNRS)-Institut Gustave Roussy (IGR)-Université Paris-Sud - Paris 11 (UP11)

Subjects
[SDV.BIO]Life Sciences [q-bio]/Biotechnology, [SDV]Life Sciences [q-bio], Sepiolite, 020101 civil engineering, 02 engineering and technology, [SDV.BBM.BM] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, 0201 civil engineering, chemistry.chemical_compound, Adsorption, Plasmid, Plasmid extraction, Geochemistry and Petrology, [CHIM]Chemical Sciences, ComputingMilieux_MISCELLANEOUS, Bacterial transformation, [PHYS]Physics [physics], Bionanohybrids, [CHIM.MATE] Chemical Sciences/Material chemistry, biology, Chemistry, Geology, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, DNA, [CHIM.MATE]Chemical Sciences/Material chemistry, 021001 nanoscience & nanotechnology, biology.organism_classification, Nanomaterial, Combinatorial chemistry, DNA extraction, [SDV.BIO] Life Sciences [q-bio]/Biotechnology, Sepiolite Bionanohybrids Nanomaterial DNA Bacterial transformation Plasmid extraction, Nanocarriers, 0210 nano-technology, Bacteria, Transformation efficiency
Abstract

International audience; Among the various clay minerals, sepiolite, which is a natural nanofibrous silicate that exhibit a poor cell toxicity, is a potential promising nanocarrier for the non-viral and stable transfer of plasmid DNA into bacteria, mammalian and human cells. We first show here that sepiolite binds to bacteria, which can be useful in decontamination protocols. In a previous research we have shown that is possible to modulate the efficiency of the absorption of different types of DNA molecules onto sepiolite, and that the DNA previously adsorbed could be recovered preserving the DNA structure and biological activity. Taking advantage of both, the sepiolite/bacteria and sepiolite/DNA interactions, we show that pre-assembly of DNA with sepiolite and incubation of bacteria with this obtained biohybrid strongly improve the transformation efficiency, in a rapid, convenient and inexpensive method that doesn't require competent cell preparation. In addition, we also show that the controlled sepiolite and DNA binding capacities can be used to purify plasmids from bacteria, representing an advantageous alternative to onerous commercial kits. All of these results open the way to the use of sepiolite-based bionanohybrids for the development of novel biological models of interest for academic and applied sciences.

Published
2020

22. Microphthalmia transcription factor expression contributes to bone marrow failure in Fanconi anemia

Author

Filippo Rosselli, Emilie Renaud, Julie Bourseguin, Alessia Oppezzo, Patrycja Pawlikowska, Intégrité du génome et cancers (IGC), Institut Gustave Roussy (IGR)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Génétique de la Radiosensibilité (LGR), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Immunology-Hematology, Stabilité Génétique et Oncogenèse (UMR 8200), and Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, DNA repair, DNA damage, MAP Kinase Signaling System, [SDV]Life Sciences [q-bio], Smad Proteins, Ascorbic Acid, Biology, Cell Line, 03 medical and health sciences, Mice, 0302 clinical medicine, Fanconi anemia, medicine, Animals, Extracellular Signal-Regulated MAP Kinases, Transcription factor, ComputingMilieux_MISCELLANEOUS, Cholecalciferol, Mice, Knockout, Microphthalmia-Associated Transcription Factor, Plant Extracts, Nicotinic Acids, Hematopoietic stem cell, General Medicine, Dehydroepiandrosterone, Bone Marrow Failure Disorders, Microphthalmia-associated transcription factor, medicine.disease, Hematopoietic Stem Cells, FANCA, Fanconi Anemia Complementation Group Proteins, Haematopoiesis, 030104 developmental biology, medicine.anatomical_structure, Fanconi Anemia, 030220 oncology & carcinogenesis, Cancer research, DNA Damage, Research Article
Abstract

Hematopoietic stem cell (HSC) attrition is considered the key event underlying progressive BM failure (BMF) in Fanconi anemia (FA), the most frequent inherited BMF disorder in humans. However, despite major advances, how the cellular, biochemical, and molecular alterations reported in FA lead to HSC exhaustion remains poorly understood. Here, we demonstrated in human and mouse cells that loss-of-function of FANCA or FANCC, products of 2 genes affecting more than 80% of FA patients worldwide, is associated with constitutive expression of the transcription factor microphthalmia (MiTF) through the cooperative, unscheduled activation of several stress-signaling pathways, including the SMAD2/3, p38 MAPK, NF-κB, and AKT cascades. We validated the unrestrained Mitf expression downstream of p38 in Fanca(–/–) mice, which display hallmarks of hematopoietic stress, including loss of HSC quiescence, DNA damage accumulation in HSCs, and reduced HSC repopulation capacity. Importantly, we demonstrated that shRNA-mediated downregulation of Mitf expression or inhibition of p38 signaling rescued HSC quiescence and prevented DNA damage accumulation. Our data support the hypothesis that HSC attrition in FA is the consequence of defects in the DNA-damage response combined with chronic activation of otherwise transiently activated signaling pathways, which jointly prevent the recovery of HSC quiescence.

Published
2020

23. Role of PARP-catalyzed ADP-ribosylation in the Crosstalk Between DNA Strand Breaks and Epigenetic Regulation

Author

Elie Matta, Haser H. Sutcu, Alexander A. Ishchenko, Réparation de l’ADN (CNRS UMR 8200), Stabilité Génétique et Oncogenèse (UMR 8200), and Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS)

Subjects
DNA damage, Poly ADP ribose polymerase, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Structural Biology, readers of ADP-ribosylated targets, Transcriptional regulation, Epigenetics, Molecular Biology, Polymerase, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, biology, Chemistry, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, programmed DNA breaks, Chromatin, Cell biology, DNA demethylation, biology.protein, regulation of transcription, DNA and RNA ADP-ribosylation, 030217 neurology & neurosurgery, DNA
Abstract

Covalent linkage of ADP-ribose units to proteins catalyzed by poly(ADP-ribose) polymerases (PARPs) plays important signaling functions in a plethora of cellular processes including DNA damage response, chromatin organization, and gene transcription. Poly- and mono-ADP-ribosylation of target macromolecules are often responsible both for the initiation and for coordination of these processes in mammalian cells. Currently, the number of cellular targets for ADP-ribosylation is rapidly expanding, and the molecular mechanisms underlying the broad substrate specificity of PARPs present enormous interest. In this review, the roles of PARP-mediated modifications of protein and nucleic acids, the readers of ADP-ribosylated structures, and the origin and function of programmed DNA strand breaks in PARP activation, transcription regulation, and DNA demethylation are discussed.

Published
2020

24. Defective transcription of ATF3 responsive genes, a marker for Cockayne Syndrome

Author

Alain Sarasin, Alexey Epanchintsev, Cathy Obringer, Nadège Calmels, Frédéric Coin, Vincent Laugel, Federico Costanzo, Marc-Alexander Rauschendorf, Jean-Marc Egly, Department of Functional Genomics and Cancer [Illkirch], Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Équipe labellisée Ligue Nationale Contre le Cancer 2014 [Illkirch], Ligue Nationale Contre le Cancer [Paris] (LNCC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA), Sanofi-Aventis Deutschland GmbH [Francfort, Allemagne], Molecular Health GmbH [Heidelberg, Allemagne], Laboratoire de Génétique Médicale (LGM), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Department of Pediatric Neurology [Strasbourg], CHU Strasbourg, Intégrité du génome et cancers (IGC), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut Gustave Roussy (IGR)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), A.E. was sponsored by the ERC and AFM grants, M-A.R. was supported by French Ministère des Affaires Etrangères and by an Inserm Young investigator fellowship and F.C. by a Marie Curie fellowship. This study was supported by the grant ANR-10-LABX-0030-INRT, a French State fund managed by the Agence Nationale de la Recherche under the frame program Investissements d’Avenir ANR-10-IDEX-0002-02., ANR-10-IDEX-0002,UNISTRA,Par-delà les frontières, l'Université de Strasbourg(2010), Bodescot, Myriam, Initiative d'excellence - Par-delà les frontières, l'Université de Strasbourg - - UNISTRA2010 - ANR-10-IDEX-0002 - IDEX - VALID, Institut Gustave Roussy (IGR)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), and Institut Gustave Roussy (IGR)

Subjects
[SDV]Life Sciences [q-bio], lcsh:Medicine, RNA polymerase II, [SDV.GEN.GH] Life Sciences [q-bio]/Genetics/Human genetics, [SDV.BBM.BM] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Cockayne syndrome, 03 medical and health sciences, 0302 clinical medicine, Transcription (biology), medicine, lcsh:Science, Gene, 030304 developmental biology, 0303 health sciences, Multidisciplinary, biology, lcsh:R, RNA, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, NIPBL, medicine.disease, Molecular biology, Gene expression profiling, [SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics, biology.protein, lcsh:Q, Immediate early gene, 030217 neurology & neurosurgery
Abstract

Cockayne syndrome (CS) is a rare genetic disorder caused by mutations (dysfunction) in CSA and CSB. CS patients exhibit mild photosensitivity and severe neurological problems. Currently, CS diagnosis is based on the inefficiency of CS cells to recover RNA synthesis upon genotoxic (UV) stress. Indeed, upon genotoxic stress, ATF3, an immediate early gene is activated to repress up to 5000 genes encompassing its responsive element for a short period of time. On the contrary in CS cells, CSA and CSB dysfunction impairs the degradation of the chromatin-bound ATF3, leading to a permanent transcriptional arrest as observed by immunofluorescence and ChIP followed by RT-PCR. We analysed ChIP-seq of Pol II and ATF3 promoter occupation analysis and RNA sequencing-based gene expression profiling in CS cells, as well as performed immunofluorescence study of ATF3 protein stability and quantitative RT-PCR screening in 64 patient cell lines. We show that the analysis of few amount (as for example CDK5RAP2, NIPBL and NRG1) of ATF3 dependent genes, could serve as prominent molecular markers to discriminate between CS and non-CS patient’s cells. Such assay can significantly simplify the timing and the complexity of the CS diagnostic procedure in comparison to the currently available methods.

Published
2020

25. Mechanisms generating cancer genome complexity: a look back at the interphase breakage model

Author

Toledo, Franck, Debatisse, Michelle, Toledo, Franck, Dynamique de l'information génétique : bases fondamentales et cancer (DIG CANCER), Institut Curie [Paris]-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Institut Curie [Paris], Institut Gustave Roussy (IGR), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), and Centre National de la Recherche Scientifique (CNRS)-Institut Curie [Paris]-Sorbonne Université (SU)

Subjects
[SDV] Life Sciences [q-bio], [SDV]Life Sciences [q-bio]
Abstract

e-letter; Understanding the mechanims responsible for cancer genome complexity has been an important goal for many decades. Umbreit et al. recently combined live cell imaging and single cell genome sequencing to analyze the cascade of genome rearrangements following the formation of a chromosome bridge in human cells (1). Their results suggest that this bridge leads to an initial breakage-fusion-bridge (BFB) cycle, followed by additional BFB cycles interwoven with episodes of micronucleation and chromothripsis, to generate complex genome rearrangements (1). This conclusion is strikingly consistent with the previously proposed “interphase breakage model” (2).

Published
2020

26. First description of ultramutated endometrial cancer caused by germline loss-of-function and somatic exonuclease domain mutations in POLE gene

Author

Victor Evangelista de Faria Ferraz, Wilson A. Silva, Sergey Nikolaev, Patricia Kannouche, Mariângela Ottoboni Brunaldi, Fernando Chahud, Reginaldo Cruz Alves Rosa, Andrey A. Yurchenko, Alfredo Ribeiro-Silva, CNRS (UMR9019), Centre National de la Recherche Scientifique (CNRS), Universidade de São Paulo (USP), Biomarqueurs prédictifs et nouvelles stratégies moléculaires en thérapeutique anticancéreuse (U981), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Intégrité du génome et cancers (IGC), Institut Gustave Roussy (IGR)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Stabilité Génétique et Oncogenèse (UMR 8200), and Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Mutation rate, Somatic cell, [SDV]Life Sciences [q-bio], Biology, QH426-470, medicine.disease_cause, Germline, Frameshift mutation, 03 medical and health sciences, 0302 clinical medicine, Endometrial cancer, medicine, Ultramutated phenotype, Genetics, Allele, Molecular Biology, Gene, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, Mutation, TMB, Wild type, CARCINOGÊNESE, 3. Good health, POLE exonuclease mutation, 030220 oncology & carcinogenesis, Targeted sequencing
Abstract

Endometrial cancer (EC) harboring heterozygous POLE proofreading inactivating mutations (POLE-exo*) is associated with an increased number of somatic mutations that result in a distinctive anti-tumor immune response. However, the consequences of such POLE mutations in the context of the missing wild-type allele have not yet been described in endometrial tumors. A 72-year-old woman harboring a germline monoallelic frameshift mutation (p.Pro269fsTer26) in POLE was diagnosed with an EC having a somatic heterozygous mutation in the exonuclease domain of POLE (S459F). Targeted gene sequencing revealed an ultramutated phenotype (381 mutations/Mb) in the tumor and a 2-fold excess of mutations on the DNA leading strand. Additionally, we observed a mutational signature similar to the COSMIC signature 10, a higher mutation rate in this tumor than in endometrial tumors with heterozygous POLE-exo*, and an increased number of T lymphocytes. This is the first report of an ultramutated EC harboring a somatic POLE-exo* mutation in association with a germline loss-of-function mutation in this gene. The absence of a wild type POLE allele led to a particularly high mutational burden.

Published
2020

27. Data on PAGE analysis and MD simulation for the interaction of endonuclease Apn1 from Saccharomyces cerevisiae with DNA substrates containing 5,6-dihydrouracyl and 2-aminopurine

Author

Vladimir V. Koval, Elena Dyakonova, Alexander A. Lomzov, Alexander A. Ishchenko, Olga S. Fedorova, Steklov Mathematical Institute [Moscow] (SMI), Russian Academy of Sciences [Moscow] (RAS), Réparation de l’ADN (CNRS UMR 8200), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Lomonosov Moscow State University (MSU), and Steklov Mathematical Institute (SMI)

Subjects
0301 basic medicine, DNA repair, Saccharomyces cerevisiae, Mutant, lcsh:Computer applications to medicine. Medical informatics, 03 medical and health sciences, chemistry.chemical_compound, Endonuclease, AP site, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], lcsh:Science (General), Multidisciplinary, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], biology, Structural gene, Wild type, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, biology.organism_classification, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], 030104 developmental biology, Biochemistry, chemistry, biology.protein, lcsh:R858-859.7, DNA, lcsh:Q1-390
Abstract

This article presents new data on nucleotide incision repair (NIR) activity of apurinic/apyrimidinic endonuclease Apn1 of Saccharomyces cerevisiae, which is known as a key player of the base excision DNA repair (BER) pathway, see "Yeast structural gene (APN1) for the major apurinic endonuclease: homology to Escherichia coli endonuclease IV" [1], "Abasic sites in DNA: repair and biological consequences in Saccharomyces cerevisiae" [2] and "Characterisation of new substrate specificities of Escherichia coli and Saccharomyces cerevisiae AP endonucleases" [3]. The characterization of NIR activity of wild type Apn1 and mutant form Ape1 H83A were made by denaturing PAGE analysis, and MD simulations of Apn1 complexed with DNA containing 5,6-dihydro-2'-deoxyuridine (DHU) and 2-aminopurine (2-aPu) residues. This data article is associated to the manuscript titled "Apurinic/apyrimidinic endonuclease Apn1 from Saccharomyces cerevisiae is recruited to the nucleotide incision repair pathway: kinetic and structural features" [4]. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Published
2018

28. Transcription-mediated organization of the replication initiation program across large genes sets common fragile sites genome-wide

Author

Anne-Marie Lachages, St eacutephane Koundrioukoff, Bernard Dutrillaux, Yan Jaszczyszyn, Michelle Debatisse, Chun-Long Chen, Viola Nähse, Sami El-Hilali, Olivier Brison, Claude Thermes, Mélanie Schmidt, Dana Azar, Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Dynamique de l'information génétique : bases fondamentales et cancer (DIG CANCER), Institut Curie [Paris]-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Université Saint-Joseph de Beyrouth (USJ), Oslo University Hospital [Oslo], Unité de Technologies Chimiques et Biologiques pour la Santé (UTCBS - UM 4 (UMR 8258 / U1022)), Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut de Systématique, Evolution, Biodiversité (ISYEB ), Muséum national d'Histoire naturelle (MNHN)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), Centre National de la Recherche Scientifique (CNRS)-Institut Curie [Paris]-Sorbonne Université (SU), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Muséum national d'Histoire naturelle (MNHN)-École pratique des hautes études (EPHE), Institut Curie-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5)-Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL (ENSCP), and Université des Antilles (UA)-Centre National de la Recherche Scientifique (CNRS)-École pratique des hautes études (EPHE)-Sorbonne Université (SU)-Muséum national d'Histoire naturelle (MNHN)

Subjects
0301 basic medicine, Genome instability, Transcription, Genetic, DNA Replication Timing, Science, General Physics and Astronomy, Replication Origin, Biology, Genome, Genomic Instability, Article, General Biochemistry, Genetics and Molecular Biology, Cell Line, S Phase, 03 medical and health sciences, 0302 clinical medicine, Transcription (biology), [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Humans, Directionality, lcsh:Science, Gene, Communication and replication, Genetics, Replication timing, Multidisciplinary, Chromosome Fragile Sites, Chromosomal fragile site, General Chemistry, 030104 developmental biology, Replication Initiation, Transcription Termination, Genetic, 030220 oncology & carcinogenesis, lcsh:Q, Fragile sites
Abstract

Common fragile sites (CFSs) are chromosome regions prone to breakage upon replication stress known to drive chromosome rearrangements during oncogenesis. Most CFSs nest in large expressed genes, suggesting that transcription could elicit their instability; however, the underlying mechanisms remain elusive. Genome-wide replication timing analyses here show that stress-induced delayed/under-replication is the hallmark of CFSs. Extensive genome-wide analyses of nascent transcripts, replication origin positioning and fork directionality reveal that 80% of CFSs nest in large transcribed domains poor in initiation events, replicated by long-travelling forks. Forks that travel long in late S phase explains CFS replication features, whereas formation of sequence-dependent fork barriers or head-on transcription–replication conflicts do not. We further show that transcription inhibition during S phase, which suppresses transcription–replication encounters and prevents origin resetting, could not rescue CFS stability. Altogether, our results show that transcription-dependent suppression of initiation events delays replication of large gene bodies, committing them to instability., Common Fragile Sites (CFSs) are chromosome regions prone to breakage upon replication stress known to drive chromosome rearrangements during oncogenesis. Here the authors use genome-wide and single cell techniques to assess how replication timing and transcriptional activity correlate with genome stability.

Published
2019

29. HIV-1 Tat protein induces DNA damage in human peripheral blood B-lymphocytes via mitochondrial ROS production

Author

Tatyana Tsfasman, Ruy A. N. Louzada, Vlada V. Zakharova, Yegor S. Vassetzky, Eric Oksenhendler, Aline Hamade, Diego Germini, Rawan El-Amine, Chrystèle Bilhou-Nabera, Boris V. Chernyak, Fadia Najjar, M. Lipinski, Eugene V. Sheval, Corinne Dupuy, Signalisation, noyaux et innovations en cancérologie (UMR8126), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Faculty of Bioengineering and Bioinformatics, Moscow, Stabilité Génétique et Oncogenèse (UMR 8200), Centre de Recherche Saint-Antoine (UMRS893), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Department of Biology, Faculty of Sciences II, Lebanese University [Beirut] (LU), Synthèse et Physico-Chimie de Molécules d'Intérêt Biologique (SPCMIB), Institut de Chimie de Toulouse (ICT-FR 2599), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Service d'Immunopathologie Clinique, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Lomonosov Moscow State University (MSU), Faculty of Bioengineering and Bioinformatics, Moscow State University, Centre de Recherche Saint-Antoine (CR Saint-Antoine), Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), and Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP)

Subjects
0301 basic medicine, Mitochondrial ROS, DNA damage, Clinical Biochemistry, Respiratory chain, [SDV.CAN]Life Sciences [q-bio]/Cancer, Biology, Mitochondrion, B-cell lymphomas, medicine.disease_cause, Biochemistry, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, medicine, Humans, lcsh:QH301-705.5, ComputingMilieux_MISCELLANEOUS, chemistry.chemical_classification, B-Lymphocytes, Reactive oxygen species, lcsh:R5-920, Organic Chemistry, NF-kappa B, Glutathione, Molecular biology, 3. Good health, Mitochondria, 030104 developmental biology, chemistry, lcsh:Biology (General), Oxidative stress, 030220 oncology & carcinogenesis, HIV-1, tat Gene Products, Human Immunodeficiency Virus, Tat, Reactive Oxygen Species, Carcinogenesis, lcsh:Medicine (General), Signal Transduction, Research Paper
Abstract

Human immunodeficiency virus (HIV) infection is associated with B-cell malignancies in patients though HIV-1 is not able to infect B-cells. The rate of B-cell lymphomas in HIV-infected individuals remains high even under the combined antiretroviral therapy (cART) that reconstitutes the immune function. Thus, the contribution of HIV-1 to B-cell oncogenesis remains enigmatic. HIV-1 induces oxidative stress and DNA damage in infected cells via multiple mechanisms, including viral Tat protein. We have detected elevated levels of reactive oxygen species (ROS) and DNA damage in B-cells of HIV-infected individuals. As Tat is present in blood of infected individuals and is able to transduce cells, we hypothesized that it could induce oxidative DNA damage in B-cells promoting genetic instability and malignant transformation. Indeed, incubation of B-cells isolated from healthy donors with purified Tat protein led to oxidative stress, a decrease in the glutathione (GSH) levels, DNA damage and appearance of chromosomal aberrations. The effects of Tat relied on its transcriptional activity and were mediated by NF-κB activation. Tat stimulated oxidative stress in B-cells mostly via mitochondrial ROS production which depended on the reverse electron flow in Complex I of respiratory chain. We propose that Tat-induced oxidative stress, DNA damage and chromosomal aberrations are novel oncogenic factors favoring B-cell lymphomas in HIV-1 infected individuals., Graphical abstract fx1, Highlights • B-cells of HIV-infected individuals exhibit elevated levels of oxidative stress, DNA damage and chromosomal aberrations. • Purified HIV-1 Tat protein reproduces this effect and induces oxidative stress and DNA damage in B-cells. • HIV-1 Tat induces mitochondrial oxidative stress and activates NF-kB in B-cells. • This condition increases the risk of developing chromosomal abnormalities and translocations.

Published
2018

30. CtIP fusion to Cas9 enhances transgene integration by homology-dependent repair

Author

Charpentier, M., Khedher, A., Menoret, S., Brion, A., Lamribet, K., Dardillac, E., Boix, C., Perrouault, L., Tesson, L., Geny, S., De Cian, A., Itier, J, Anegon, I., Lopez, B., Giovannangeli, C., Concordet, J., Structure et Instabilité des Génomes (STRING), Muséum national d'Histoire naturelle (MNHN)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Translational Sciences [Paris] (SANOFI), SANOFI Recherche, Genetic and Cellular Engineering in Immunology and Regenerative Medicine (Team 2 - U1064 Inserm - CRTI), Centre de Recherche en Transplantation et Immunologie (U1064 Inserm - CRTI), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), ANRT, ANR-11-INBS-0014,TEFOR,Transgenèse pour les Etudes Fonctionnelles sur les Organismes modèles(2011), ANR-16-CE18-0012,STaHR,Stimulation de la Recombinaison Homologue pour la Thérapie Génique(2016), ANR-14-LAB5-0008,SOURIRAT,Nouveaux outils pour la création de rongeurs génétiquement modifiés(2014), ANR-11-LABX-0016,IGO,Immunothérapies Grand Ouest(2011), ANR-10-IBHU-0005,CESTI (TSI-IHU),Centre Européen des Sciences de la Transplantation et de l'Immunothérapie (TSI-IHU)(2010), Le Bihan, Sylvie, Infrastructures - Transgenèse pour les Etudes Fonctionnelles sur les Organismes modèles - - TEFOR2011 - ANR-11-INBS-0014 - INBS - VALID, Stimulation de la Recombinaison Homologue pour la Thérapie Génique - - STaHR2016 - ANR-16-CE18-0012 - AAPG2016 - VALID, Laboratoires communs organismes de recherche publics – PME/ETI - Nouveaux outils pour la création de rongeurs génétiquement modifiés - - SOURIRAT2014 - ANR-14-LAB5-0008 - LABCOM - VALID, Laboratoires d'excellence - Immunothérapies Grand Ouest - - IGO2011 - ANR-11-LABX-0016 - LABX - VALID, and Instituts Hospitalo-Universitaires B - Centre Européen des Sciences de la Transplantation et de l'Immunothérapie (TSI-IHU) - - CESTI (TSI-IHU)2010 - ANR-10-IBHU-0005 - IBHU - VALID

Subjects
[SDV.MHEP] Life Sciences [q-bio]/Human health and pathology, Science, lcsh:Q, lcsh:Science, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract

International audience; In genome editing with CRISPR-Cas9, transgene integration often remains challenging. Here, we present an approach for increasing the efficiency of transgene integration by homology-dependent repair (HDR). CtIP, a key protein in early steps of homologous recombination, is fused to Cas9 and stimulates transgene integration by HDR at the human AAVS1 safe harbor locus. A minimal N-terminal fragment of CtIP, designated HE for HDR enhancer, is sufficient to stimulate HDR and this depends on CDK phosphorylation sites and the multimerization domain essential for CtIP activity in homologous recombination. HDR stimulation by Cas9-HE, however, depends on the guide RNA used, a limitation that may be overcome by testing multiple guides to the locus of interest. The Cas9-HE fusion is simple to use and allows obtaining twofold or more efficient transgene integration than that with Cas9 in several experimental systems, including human cell lines, iPS cells, and rat zygotes.

Published
2018

31. Xeroderma pigmentosum in South Africa: Evidence for a prevalent founder effect

Author

Hamid Reza Rezvani, Fanny Morice-Picard, Frédéric Austerlitz, Alain Taieb, François Cartault, Alain Sarasin, Cécile Ged, M. Kgokolo, Mike Sathekge, Eco-Anthropologie et Ethnobiologie (EAE), Muséum national d'Histoire naturelle (MNHN)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Biologie et écologie tropicale et méditerranéenne [2007-2010] (BETM), Université de Perpignan Via Domitia (UPVD)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS), Génomes et cancer (GC (FRE2939)), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Stabilité Génétique et Oncogenèse (UMR 8200), Éco-Anthropologie (EA), Biologie et écologie tropicale et méditerranéenne (BETM), and Université de Perpignan Via Domitia (UPVD)-École pratique des hautes études (EPHE)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Male, medicine.medical_specialty, Skin Neoplasms, Xeroderma pigmentosum, Adolescent, DNA Mutational Analysis, Dermatology, South Africa, Young Adult, 030207 dermatology & venereal diseases, 03 medical and health sciences, 0302 clinical medicine, medicine, Humans, Child, ComputingMilieux_MISCELLANEOUS, Xeroderma Pigmentosum Group D Protein, 030304 developmental biology, Xeroderma Pigmentosum, 0303 health sciences, [SDV.GEN.GPO]Life Sciences [q-bio]/Genetics/Populations and Evolution [q-bio.PE], business.industry, medicine.disease, Founder Effect, DNA-Binding Proteins, Child, Preschool, Mutation, Female, business, Founder effect
Abstract

International audience

Published
2019

32. DUOX2 Mutations Are Associated With Congenital Hypothyroidism With Ectopic Thyroid Gland

Author

Corinne Dupuy, Denise P. Carvalho, Magnus R. Dias-da-Silva, Jessica R Jara, Miguel Mitne-Neto, Rui M. B. Maciel, Suzana Nesi-França, Gilberto K. Furuzawa, Marina M. L. Kizys, Ruy A. N. Louzada, Institut Gustave Roussy (IGR), Stabilité Génétique et Oncogenèse (UMR 8200), and Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Male, 0301 basic medicine, medicine.medical_specialty, Candidate gene, Endocrinology, Diabetes and Metabolism, DNA Mutational Analysis, Clinical Biochemistry, Thyroid Gland, Context (language use), medicine.disease_cause, Biochemistry, Thyroid dysgenesis, Cohort Studies, 03 medical and health sciences, symbols.namesake, Endocrinology, Protein Domains, Internal medicine, Congenital Hypothyroidism, medicine, Humans, Genetic Predisposition to Disease, ComputingMilieux_MISCELLANEOUS, Genetic Association Studies, Congenital Hypothyroidism with Ectopic Thyroid, Sanger sequencing, Mutation, business.industry, Biochemistry (medical), Thyroid, Infant, Newborn, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, medicine.disease, Dual Oxidases, 3. Good health, Congenital hypothyroidism, HEK293 Cells, 030104 developmental biology, medicine.anatomical_structure, Thyroid Dysgenesis, symbols, Female, business
Abstract

Context Thyroid dysgenesis (TD) is the leading cause of congenital hypothyroidism (CH). The etiology of TD remains unknown in ∼90% of cases, the most common form being thyroid ectopia (TE) (48% to 61%). Objective To search for candidate genes in hypothyroid children with TE. Design, Setting, and Participants We followed a cohort of 268 children with TD and performed whole-exome sequencing (WES) in three children with CH with TE (CHTE) and compared them with 18 thyroid-healthy controls. We then screened an additional 41 children with CHTE by Sanger sequencing and correlated the WES and Sanger molecular findings with in vitro functional analysis. Main Outcome Measures Genotyping, mutation prediction analysis, and in vitro functional analysis. Results We identified seven variants in the DUOX2 gene, namely G201E, L264CfsX57, P609S, M650T, E810X, M822V, and E1017G, and eight known variations. All children carrying DUOX2 variations had high thyroid-stimulating hormone levels at neonatal diagnosis. All mutations were localized in the N-terminal segment, and three of them led to effects on cell surface targeting and reactive oxygen species generation. The DUOX2 mutants also altered the interaction with the maturation factor DUOXA2 and the formation of a stable DUOX2/DUOXA2 complex at the cell surface, thereby impairing functional enzymatic activity. We observed no mutations in the classic genes related to TD or in the DUOX1 gene. Conclusion Our findings suggest that, in addition to thyroid hormonogenesis, the DUOX2 N-terminal domain may play a role in thyroid development.

Published
2017

33. Aberrant base excision repair pathway of oxidatively damaged DNA: Implications for degenerative diseases

Author

Dmitry O. Zharkov, Murat Saparbaev, Bakhyt T. Matkarimov, Ibtissam Talhaoui, Thierry Tchénio, Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), National Laboratory Astana, Nazarbayev University [Kazakhstan], and Novosibirsk State University (NSU)

Subjects
0301 basic medicine, DNA Repair, DNA repair, [SDV]Life Sciences [q-bio], Biology, Biochemistry, MBD4, 03 medical and health sciences, 0302 clinical medicine, Neoplasms, Physiology (medical), Animals, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Replication protein A, ComputingMilieux_MISCELLANEOUS, Cellular Senescence, Neurodegenerative Diseases, DNA, Base excision repair, Molecular biology, 3. Good health, Very short patch repair, Oxidative Stress, 030104 developmental biology, DNA glycosylase, 030220 oncology & carcinogenesis, DNA mismatch repair, Oxidation-Reduction, DNA Damage, Nucleotide excision repair
Abstract

In cellular organisms composition of DNA is constrained to only four nucleobases A, G, T and C, except for minor DNA base modifications such as methylation which serves for defence against foreign DNA or gene expression regulation. Interestingly, this severe evolutionary constraint among other things demands DNA repair systems to discriminate between regular and modified bases. DNA glycosylases specifically recognize and excise damaged bases among vast majority of regular bases in the base excision repair (BER) pathway. However, the mismatched base pairs in DNA can occur from a spontaneous conversion of 5-methylcytosine to thymine and DNA polymerase errors during replication. To counteract these mutagenic threats to genome stability, cells evolved special DNA repair systems that target the non-damaged DNA strand in a duplex to remove mismatched regular DNA bases. Mismatch-specific adenine- and thymine-DNA glycosylases (MutY/MUTYH and TDG/MBD4, respectively) initiated BER and mismatch repair (MMR) pathways can recognize and remove normal DNA bases in mismatched DNA duplexes. Importantly, in DNA repair deficient cells bacterial MutY, human TDG and mammalian MMR can act in the aberrant manner: MutY and TDG removes adenine and thymine opposite misincorporated 8-oxoguanine and damaged adenine, respectively, whereas MMR removes thymine opposite to O6-methylguanine. These unusual activities lead either to mutations or futile DNA repair, thus indicating that the DNA repair pathways which target non-damaged DNA strand can act in aberrant manner and introduce genome instability in the presence of unrepaired DNA lesions. Evidences accumulated showing that in addition to the accumulation of oxidatively damaged DNA in cells, the aberrant DNA repair can also contribute to cancer, brain disorders and premature senescence. For example, the aberrant BER and MMR pathways for oxidized guanine residues can lead to trinucleotide expansion that underlies Huntington's disease, a severe hereditary neurodegenerative syndrome. This review summarises the present knowledge about the aberrant DNA repair pathways for oxidized base modifications and their possible role in age-related diseases.

Published
2017

34. NADPH Oxidase NOX4 Is a Critical Mediator of BRAFV600E-Induced Downregulation of the Sodium/Iodide Symporter in Papillary Thyroid Carcinomas

Author

Abderrahmane Al Bouzidi, Corinne Dupuy, Jérémy Cailloux, Juliana Cazarin, Jennifer R. Cracchiolo, James A. Fagin, Rabii Ameziane-El-Hassani, Aurore Carré, Martin Schlumberger, Jeffrey A. Knauf, Abir Al Ghuzlan, Mohammed El Mzibri, Naima Azouzi, Dana M. Hartl, Michel Polak, Abdelkarim Filali-Maltouf, Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Pathologie morphologique, Département de biologie et pathologie médicales [Gustave Roussy], Institut Gustave Roussy (IGR)-Institut Gustave Roussy (IGR), LPP - Laboratoire de Phonétique et Phonologie - UMR 7018 (LPP), Université Sorbonne Nouvelle - Paris 3-Centre National de la Recherche Scientifique (CNRS), Génétique des maladies multifactorielles (GMM), Université de Lille, Droit et Santé-Centre National de la Recherche Scientifique (CNRS), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Biology and Mediacl Research Unit, Centre National de l'Energie, des Sciences et Techniques Nucléaires, Faculté des sciences, Laboratoire de microbiologie et biologie moléculaire, Université Mohammed V de Rabat [Agdal], Médecine nucléaire, Département d'imagerie médicale [Gustave Roussy], Institut Gustave Roussy (IGR), and Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Sodium-iodide symporter, medicine.medical_specialty, endocrine system diseases, Physiology, Clinical Biochemistry, [SDV.CAN]Life Sciences [q-bio]/Cancer, Biochemistry, Papillary thyroid cancer, Thyroid carcinoma, 03 medical and health sciences, 0302 clinical medicine, Downregulation and upregulation, Internal medicine, medicine, Molecular Biology, ComputingMilieux_MISCELLANEOUS, General Environmental Science, NADPH oxidase, biology, Chemistry, Thyroid, NOX4, Cell Biology, medicine.disease, 030104 developmental biology, Endocrinology, medicine.anatomical_structure, 030220 oncology & carcinogenesis, Symporter, biology.protein, Cancer research, General Earth and Planetary Sciences
Abstract

Aims: The BRAFV600E oncogene, reported in 40%–60% of papillary thyroid cancer (PTC), has an important role in the pathogenesis of PTC. It is associated with the loss of thyroid iodide-metabolizing genes, such as sodium/iodide symporter (NIS), and therefore with radioiodine refractoriness. Inhibition of mitogen-activated protein kinase (MAPK) pathway, constitutively activated by BRAFV600E, is not always efficient in resistant tumors suggesting that other compensatory mechanisms contribute to a BRAFV600E adaptive resistance. Recent studies pointed to a key role of transforming growth factor β (TGF-β) in BRAFV600E-induced effects. The reactive oxygen species (ROS)-generating NADPH oxidase NOX4, which is increased in PTC, has been identified as a new key effector of TGF-β in cancer, suggestive of a potential role in BRAFV600E-induced thyroid tumors. Results: Here, using two human BRAFV600E-mutated thyroid cell lines and a rat thyroid cell line expressing BRAFV600E in a conditional manner, we show tha...

Published
2017

35. The FANCM family Mph1 helicase localizes to the mitochondria and contributes to mtDNA stability

Author

Michael Lisby, Xuejiao Yang, Gerard Mazón, Manuel Bernal, Stabilité Génétique et Oncogenèse (UMR 8200), and Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Mitochondrial DNA, Saccharomyces cerevisiae Proteins, DNA repair, [SDV]Life Sciences [q-bio], Respiratory chain, Saccharomyces cerevisiae, Biochemistry, Genome, DNA, Mitochondrial, DEAD-box RNA Helicases, 03 medical and health sciences, 0302 clinical medicine, FANCM, Molecular Biology, 030304 developmental biology, 0303 health sciences, biology, Helicase, Cell Biology, Nuclear DNA, Cell biology, Mitochondria, Protein Transport, Mitochondrial DNA repair, 030220 oncology & carcinogenesis, biology.protein, Oxidation-Reduction, DNA Damage
Abstract

Mitochondria are membrane-bound organelles found in eukaryotic cells where they generate energy through the respiratory chain. They contain their own genome that encodes genes critical to the mitochondrial function, but most of their protein content is synthetized from nuclear encoded genes. Damages to the mtDNA can cause mutations and rearrangements with an impact on the respiratory functions of the cell. DNA repair factors are able to localize to mitochondria to restore mtDNA integrity and ensure its proper inheritance. We describe in this article the mitochondrial localization of the Mph1/FANCM helicase that serves critical roles in nuclear DNA repair processes. Mph1 localizes to mitochondria and its functions contribute to the mtDNA integrity under mtDNA damaging conditions.

Published
2019

36. Mécanisme de stimulation de la liaison à l'ADN des facteurs de transcription par l'endonucléase apurinique/apyrimidinique humaine 1, APE1

Author

Eric Le Cam, Murat Saparbaev, Anton V. Endutkin, Milena Bazlekowa-Karaban, Paulina Prorok, Zhiger Akishev, Alexander V. Popov, Amangeldy K. Bissenbaev, Bakhyt T. Matkarimov, Sabira Taipakova, Regina Groisman, Sonia Baconnais, Barbara Tudek, Dmitry O. Zharkov, Dominika Zembrzuska, Alexander A. Ishchenko, Institut de génétique humaine (IGH), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Maintenance des génomes, Microscopies Moléculaire et Bionanosciences, Centre National de la Recherche Scientifique-Université Paris Sud, Maintenance des génomes, Department of Molecular Biology and Genetics, Al-Farabi Kazakh National University, European Synchrotron Radiation Facility (ESRF), Laboratoire Epigenetique et Cancer, Centre National de la Recherche Scientifique (CNRS), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), National Laboratory Astana, Nazarbayev University [Kazakhstan], Signalisation, noyaux et innovations en cancérologie (UMR8126), Novosibirsk State University (NSU), Institute of Biochemistry and Biophysics, and Polska Akademia Nauk = Polish Academy of Sciences (PAN)

Subjects
Models, Molecular, DNA damage, [SDV]Life Sciences [q-bio], oxidative damage, Biochemistry, base excision repair, AP endonuclease, redox regulation, 03 medical and health sciences, Endonuclease, chemistry.chemical_compound, 0302 clinical medicine, transcription factors, DNA-(Apurinic or Apyrimidinic Site) Lyase, Humans, oxidative stress, AP site, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Amino Acid Sequence, Protein Structure, Quaternary, Molecular Biology, Transcription factor, nucleotide incision repair, 030304 developmental biology, 0303 health sciences, biology, Cooperative binding, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, DNA, Cell Biology, Base excision repair, Dna damage, chemistry, 030220 oncology & carcinogenesis, Biocatalysis, biology.protein, Biophysics, Protein Multimerization, Protein Binding
Abstract

International audience; Aerobic respiration generates reactive oxygen species (ROS), which can damage nucleic acids, proteins and lipids. A number of transcription factors (TFs) contain redox-sensitive cysteine residues at their DNA-binding sites, hence ROS-induced thiol oxidation strongly inhibits their recognition of the cognate DNA sequences. Major human apurinic/apyrimidinic (AP) endonuclease 1 (APE1/APEX1/HAP-1), referred also as a redox factor 1 (Ref-1), stimulates the DNA binding activities of the oxidized TFs such as AP-1 and NF-κB. Also, APE1 participates in the base excision repair (BER) and nucleotide incision repair (NIR) pathways to remove oxidative DNA base damage. At present, the molecular mechanism underlying the TF-stimulating/redox function of APE1 and its biological role remains disputed. Here, we provide evidence that, instead of direct cysteine reduction in TFs by APE1, APE1-catalyzed NIR and TF-stimulating activities may be based on transient cooperative binding of APE1 to DNA and induction of conformational changes in the helix. The structure of DNA duplex strongly influences NIR and TF-stimulating activities. Homologous plant AP endonucleases lacking conserved cysteine residues stimulate DNA binding of the p50 subunit of NF-κB. APE1 acts synergistically with low-molecular-weight reducing agents on TFs. Finally, APE1 stimulates DNA binding of the redox-insensitive p50-C62S mutant protein. Electron microscopy imaging of APE1 complexes with DNA revealed preferential polymerization of APE1 on the gapped and intrinsically curved DNA duplexes. Molecular modeling offers a structural explanation how full-length APE1 can oligomerize on DNA. In conclusion, we propose that DNA-directed APE1 oligomerization can be regarded as a substitute for diffusion of APE1 along the DNA contour to probe for anisotropic flexibility. APE1 oligomers exacerbate pre-existing distortions in DNA and enable both NIR activity and DNA binding by TFs regardless of their oxidation state.; La respiration aérobie génère des espèces réactives de l'oxygène (ERO), qui peuvent endommager les acides nucléiques, les protéines et les lipides. Un certain nombre de facteurs de transcription (TF) contiennent des résidus de cystéine sensibles à l'oxydoréduction sur leurs sites de liaison à l'ADN, c'est pourquoi l'oxydation des thiols induite par les ROS inhibe fortement leur reconnaissance des séquences d'ADN apparentées. L'endonucléase 1 apurinique /apyrimidinique (AP) humaine majeure (APE1/APEX1/HAP-1), également appelée facteur redox 1 (Ref-1), stimule les activités de liaison à l'ADN des TF oxydés tels que AP-1 et NF-κB. De plus, l'APE1 participe aux voies de réparation par excision des bases (BER) et de réparation par incision des nucléotides (NIR) pour éliminer les dommages oxydatifs des bases de l'ADN. À l'heure actuelle, le mécanisme moléculaire qui sous-tend la fonction de stimulation des TF/redox de l'APE1 et son rôle biologique restent contestés. Ici, nous apportons la preuve qu'au lieu d'une réduction directe de la cystéine dans les TF par l'APE1, les activités NIR et de stimulation des TF catalysées par l'APE1 peuvent être basées sur une liaison coopérative transitoire de l'APE1 à l'ADN et l'induction de changements conformationnels dans l'hélice. La structure du duplex de l'ADN influence fortement les activités de stimulation de la RNI et de la TF. Les endonucléases AP des plantes homologues dépourvues de résidus de cystéine conservés stimulent la liaison de l'ADN de la sous-unité p50 de NF-κB. L'APE1 agit en synergie avec des agents réducteurs de faible poids moléculaire sur les TF. Enfin, APE1 stimule la liaison de l'ADN de la protéine mutante p50-C62S insensible à l'oxydoréduction. L'imagerie au microscope électronique des complexes APE1 avec l'ADN a révélé une polymérisation préférentielle de l'APE1 sur les duplex d'ADN à espacement et à courbure intrinsèque. La modélisation moléculaire offre une explication structurelle de la façon dont l'APE1 peut s'oligomériser sur l'ADN. En conclusion, nous proposons que l'oligomérisation de l'APE1 dirigée par l'ADN peut être considérée comme un substitut à la diffusion de l'APE1 le long du contour de l'ADN pour sonder la flexibilité anisotrope. Les oligomères de l'APE1 exacerbent les distorsions préexistantes de l'ADN et permettent à la fois une activité NIR et une liaison de l'ADN par les TF, quel que soit leur état d'oxydation. Traduit avec www.DeepL.com/Translator (version gratuite)

Published
2019

37. Genetic susceptibility to radiation-related differentiated thyroid cancers: a systematic review of literature

Author

Fabienne Lesueur, Martin Schlumberger, Florent de Vathaire, Jean-Baptiste Cazier, Sylvie Chevillard, Anne Boland, Corinne Dupuy, Catherine Ory, Monia Zidane, Jean-François Deleuze, Nadia Haddy, University of Birmingham [Birmingham], Laboratoire de Cancérologie Expérimentale (LCE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Médecine nucléaire, Département d'imagerie médicale [Gustave Roussy], Institut Gustave Roussy (IGR)-Institut Gustave Roussy (IGR), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Fondation Jean Dausset CEPH, Centre National de Genotypage, Centre de recherche en épidémiologie et santé des populations (CESP), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Sud - Paris 11 (UP11)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Paul Brousse-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre National de Génotypage (CNG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), and Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris-Sud - Paris 11 (UP11)-Hôpital Paul Brousse-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)

Subjects
0301 basic medicine, Cancer Research, medicine.medical_specialty, Neoplasms, Radiation-Induced, DNA Repair, Transcription, Genetic, DNA damage, DNA repair, Endocrinology, Diabetes and Metabolism, medicine.medical_treatment, Population, Thyroid Gland, [SDV.CAN]Life Sciences [q-bio]/Cancer, Bioinformatics, Polymorphism, Single Nucleotide, 03 medical and health sciences, Direct DNA damage, 0302 clinical medicine, Endocrinology, Molecular genetics, medicine, Genetic predisposition, Humans, Genetic Predisposition to Disease, Thyroid Neoplasms, education, Thyroid cancer, ComputingMilieux_MISCELLANEOUS, education.field_of_study, business.industry, medicine.disease, 3. Good health, Radiation therapy, 030104 developmental biology, Oncology, 030220 oncology & carcinogenesis, business, Signal Transduction
Abstract

The first study establishing exposure to ionizing radiations (IRs) as a risk factor for differentiated thyroid cancer (DTC) was published 70 years ago. Given that radiation exposure causes direct DNA damage, genetic alterations in the different DNA repair mechanisms are assumed to play an important role in long-term IR-induced DNA damage prevention. Individual variations in DNA repair capacity may cause different reactions to damage made by IR exposure. The aim of this review is to recapitulate current knowledge about constitutional genetic polymorphisms found to be significantly associated with DTC occurring after IR exposure. Studies were screened online using electronic databases – only fully available articles, and studies performed among irradiated population or taking radiation exposure as adjustment factors and showing significant results are included. Nine articles were identified. Ten variants in/near to genes in six biological pathways, namely thyroid activity regulations, generic transcription, RET signaling, ATM signaling and DNA repair pathways were found to be associated with radiation-related DTC in these studies. Only seven variants were found to be in interaction with IR exposure in DTC risk. Most of these variants are also associated to sporadic DTC and are not specific to IR-related DTC. In the published studies, no data on children treated with radiotherapy is described. In conclusion, more studies carried out on larger cohorts or on case–control studies with well-documented individual radiation dose estimations are needed to get a comprehensive picture of genetic susceptibility factors involved in radiation-related DTC.

Published
2019

38. Mechanism of stimulation of DNA binding of the transcription factors by human apurinic/apyrimidinic endonuclease 1, APE1

Author

Bazlekowa-Karaban, Milena, Prorok, Paulina, Baconnais, Sonia, Taipakova, Sabira, Akishev, Zhiger, Zembrzuska, Dominika, Popov, Alexander, Endutkin, Anton, Groisman, Regina, Ishchenko, Alexander, Matkarimov, Bakhyt, Bissenbaev, Amangeldy, Le Cam, Eric, Zharkov, Dmitry, Tudek, Barbara, Saparbaev, Murat, Institut de génétique humaine (IGH), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Maintenance des génomes, Microscopies Moléculaire et Bionanosciences, Centre National de la Recherche Scientifique-Université Paris Sud, Maintenance des génomes, Department of Molecular Biology and Genetics, Al-Farabi Kazakh National University, European Synchrotron Radiation Facility (ESRF), Laboratoire Epigenetique et Cancer, Centre National de la Recherche Scientifique (CNRS), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), National Laboratory Astana, Nazarbayev University [Kazakhstan], Signalisation, noyaux et innovations en cancérologie (UMR8126), Novosibirsk State University (NSU), Institute of Biochemistry and Biophysics, and Polska Akademia Nauk = Polish Academy of Sciences (PAN)

Subjects
redox regulation, AP endonuclease, transcription factors, [SDV]Life Sciences [q-bio], oxidative stress, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, oxidative damage, Dna damage, base excision repair, nucleotide incision repair
Abstract

International audience; Aerobic respiration generates reactive oxygen species (ROS), which can damage nucleic acids, proteins and lipids. A number of transcription factors (TFs) contain redox-sensitive cysteine residues at their DNA-binding sites, hence ROS-induced thiol oxidation strongly inhibits their recognition of the cognate DNA sequences. Major human apurinic/apyrimidinic (AP) endonuclease 1 (APE1/APEX1/HAP-1), referred also as a redox factor 1 (Ref-1), stimulates the DNA binding activities of the oxidized TFs such as AP-1 and NF-κB. Also, APE1 participates in the base excision repair (BER) and nucleotide incision repair (NIR) pathways to remove oxidative DNA base damage. At present, the molecular mechanism underlying the TF-stimulating/redox function of APE1 and its biological role remains disputed. Here, we provide evidence that, instead of direct cysteine reduction in TFs by APE1, APE1-catalyzed NIR and TF-stimulating activities may be based on transient cooperative binding of APE1 to DNA and induction of conformational changes in the helix. The structure of DNA duplex strongly influences NIR and TF-stimulating activities. Homologous plant AP endonucleases lacking conserved cysteine residues stimulate DNA binding of the p50 subunit of NF-κB. APE1 acts synergistically with low-molecular-weight reducing agents on TFs. Finally, APE1 stimulates DNA binding of the redox-insensitive p50-C62S mutant protein. Electron microscopy imaging of APE1 complexes with DNA revealed preferential polymerization of APE1 on the gapped and intrinsically curved DNA duplexes. Molecular modeling offers a structural explanation how full-length APE1 can oligomerize on DNA. In conclusion, we propose that DNA-directed APE1 oligomerization can be regarded as a substitute for diffusion of APE1 along the DNA contour to probe for anisotropic flexibility. APE1 oligomers exacerbate pre-existing distortions in DNA and enable both NIR activity and DNA binding by TFs regardless of their oxidation state.; La respiration aérobie génère des espèces réactives de l'oxygène (ERO), qui peuvent endommager les acides nucléiques, les protéines et les lipides. Un certain nombre de facteurs de transcription (TF) contiennent des résidus de cystéine sensibles à l'oxydoréduction sur leurs sites de liaison à l'ADN, c'est pourquoi l'oxydation des thiols induite par les ROS inhibe fortement leur reconnaissance des séquences d'ADN apparentées. L'endonucléase 1 apurinique /apyrimidinique (AP) humaine majeure (APE1/APEX1/HAP-1), également appelée facteur redox 1 (Ref-1), stimule les activités de liaison à l'ADN des TF oxydés tels que AP-1 et NF-κB. De plus, l'APE1 participe aux voies de réparation par excision des bases (BER) et de réparation par incision des nucléotides (NIR) pour éliminer les dommages oxydatifs des bases de l'ADN. À l'heure actuelle, le mécanisme moléculaire qui sous-tend la fonction de stimulation des TF/redox de l'APE1 et son rôle biologique restent contestés. Ici, nous apportons la preuve qu'au lieu d'une réduction directe de la cystéine dans les TF par l'APE1, les activités NIR et de stimulation des TF catalysées par l'APE1 peuvent être basées sur une liaison coopérative transitoire de l'APE1 à l'ADN et l'induction de changements conformationnels dans l'hélice. La structure du duplex de l'ADN influence fortement les activités de stimulation de la RNI et de la TF. Les endonucléases AP des plantes homologues dépourvues de résidus de cystéine conservés stimulent la liaison de l'ADN de la sous-unité p50 de NF-κB. L'APE1 agit en synergie avec des agents réducteurs de faible poids moléculaire sur les TF. Enfin, APE1 stimule la liaison de l'ADN de la protéine mutante p50-C62S insensible à l'oxydoréduction. L'imagerie au microscope électronique des complexes APE1 avec l'ADN a révélé une polymérisation préférentielle de l'APE1 sur les duplex d'ADN à espacement et à courbure intrinsèque. La modélisation moléculaire offre une explication structurelle de la façon dont l'APE1 peut s'oligomériser sur l'ADN. En conclusion, nous proposons que l'oligomérisation de l'APE1 dirigée par l'ADN peut être considérée comme un substitut à la diffusion de l'APE1 le long du contour de l'ADN pour sonder la flexibilité anisotrope. Les oligomères de l'APE1 exacerbent les distorsions préexistantes de l'ADN et permettent à la fois une activité NIR et une liaison de l'ADN par les TF, quel que soit leur état d'oxydation. Traduit avec www.DeepL.com/Translator (version gratuite)

Published
2019

39. Redox modifications of cysteine-containing proteins, cell cycle arrest and translation inhibition: Involvement in vitamin C-induced breast cancer cell death

Author

Stéphan Vagner, Corinne Dupuy, Amélie Heneman-Masurel, Laurence Vernis, Meng-Er Huang, Jean-Michel Camadro, Sylvain Martineau, Elie Hatem, Nadine El Banna, Camille Garcia, Thibaut Léger, Dorothée Baïlle, Stress génotoxiques et cancer, Université Paris-Sud - Paris 11 (UP11)-Institut Curie-Centre National de la Recherche Scientifique (CNRS), Institut Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Departement /u563 : Oncogenèse, Signalisation et Innovation thérapeutique, Centre de Physiopathologie Toulouse Purpan ex IFR 30 et IFR 150 (CPTP), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut Claudius Regaud, Institut de Biologie Intégrative de la Cellule (I2BC), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay, Institut Gustave Roussy (IGR), Institut Claudius Regaud, CRLCC Institut Claudius Regaud, Intégrité du génome, ARN et cancer, Institut Curie [Paris]-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Institut Curie [Paris]-Université Paris-Sud - Paris 11 (UP11), and Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP)

Subjects
0301 basic medicine, [SDV]Life Sciences [q-bio], Clinical Biochemistry, medicine.disease_cause, Biochemistry, Antioxidants, chemistry.chemical_compound, Breast cancer, 0302 clinical medicine, Vitamin C, Cytotoxicity, lcsh:QH301-705.5, ComputingMilieux_MISCELLANEOUS, Dehydroascorbic acid, lcsh:R5-920, Chemistry, Glutathione, 3. Good health, Ascorbic acid, lcsh:Medicine (General), Oxidation-Reduction, Intracellular, Research Paper, [SDV.CAN]Life Sciences [q-bio]/Cancer, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Peroxiredoxin 1, Cell Line, 03 medical and health sciences, medicine, Humans, Cysteine, Organic Chemistry, Computational Biology, Endothelial Cells, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Redoxome, Cell Cycle Checkpoints, Hydrogen Peroxide, Peroxiredoxins, 030104 developmental biology, lcsh:Biology (General), Oxidative stress, Cell culture, Protein Biosynthesis, Reactive Oxygen Species, 030217 neurology & neurosurgery
Abstract

Vitamin C (VitC) possesses pro-oxidant properties at high pharmacologic concentrations which favor repurposing VitC as an anti-cancer therapeutic agent. However, redox-based anticancer properties of VitC are yet partially understood. We examined the difference between the reduced and oxidized forms of VitC, ascorbic acid (AA) and dehydroascorbic acid (DHA), in terms of cytotoxicity and redox mechanisms toward breast cancer cells. Our data showed that AA displayed higher cytotoxicity towards triple-negative breast cancer (TNBC) cell lines in vitro than DHA. AA exhibited a similar cytotoxicity on non-TNBC cells, while only a minor detrimental effect on noncancerous cells. Using MDA-MB-231, a representative TNBC cell line, we observed that AA- and DHA-induced cytotoxicity were linked to cellular redox-state alterations. Hydrogen peroxide (H2O2) accumulation in the extracellular medium and in different intracellular compartments, and to a lesser degree, intracellular glutathione oxidation, played a key role in AA-induced cytotoxicity. In contrast, DHA affected glutathione oxidation and had less cytotoxicity. A “redoxome” approach revealed that AA treatment altered the redox state of key antioxidants and a number of cysteine-containing proteins including many nucleic acid binding proteins and proteins involved in RNA and DNA metabolisms and in energetic processes. We showed that cell cycle arrest and translation inhibition were associated with AA-induced cytotoxicity. Finally, bioinformatics analysis and biological experiments identified that peroxiredoxin 1 (PRDX1) expression levels correlated with AA differential cytotoxicity in breast cancer cells, suggesting a potential predictive value of PRDX1. This study provides insight into the redox-based mechanisms of VitC anticancer activity, indicating that pharmacologic doses of VitC and VitC-based rational drug combinations could be novel therapeutic opportunities for triple-negative breast cancer., Graphical abstract Image 1, Highlights • Ascorbic acid (AA) displays high cytotoxicity towards breast cancer cells in vitro. • H2O2 is the main mediator of AA cytotoxicity toward breast cancer cells. • AA treatment alters the redox state of large number of cysteine-containing proteins. • AA treatment triggers cell cycle arrest and translation inhibition. • PRDX1 expression levels could be a predictive biomarker for cellular response to AA.

Published
2019

40. Whole exome sequencing identifies a new mutation in the SLC19A2 gene leading to thiamine‐responsive megaloblastic anemia in an Egyptian family

Author

Ghada El-Kamah, Patrycja Pawlikowska, Said Aoufouchi, Khalda Amr, Filippo Rosselli, Medical Molecular Genetics Department, Human Genetics & Genome Research Division, National Research Center, Cairo, Egypt, Immunology-Hematology, Stabilité Génétique et Oncogenèse (UMR 8200), and Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Male, Microcephaly, Anemia, Megaloblastic, [SDV]Life Sciences [q-bio], 030105 genetics & heredity, Consanguinity, Medicine, Child, ComputingMilieux_MISCELLANEOUS, Genetics (clinical), Exome sequencing, Sanger sequencing, Genetics, biology, diabetes, Thiamine Deficiency, 3. Good health, Pedigree, SLC19A2, symbols, Thiamine, Egypt, Original Article, lcsh:QH426-470, Hearing Loss, Sensorineural, THTR‐1, 03 medical and health sciences, symbols.namesake, deafness, Exome Sequencing, Thiamine transporter, Diabetes Mellitus, Humans, Family, Megaloblastic anemia, Molecular Biology, business.industry, Bone marrow failure, Membrane Transport Proteins, Original Articles, medicine.disease, lcsh:Genetics, 030104 developmental biology, Mutation, biology.protein, bone marrow failure, business, thiamine‐responsive megaloblastic anemia
Abstract

Background The Solute Carrier Family 19 Member 2 (SLC19A2, OMIM *603941) encodes the thiamine transporter 1 (THTR‐1) that brings thiamine (Vitamin B1) into cells. THTR‐1 is the only thiamine transporter expressed in bone marrow, cochlear, and pancreatic beta cells. THTR‐1 loss‐of‐function leads to the rare recessive genetic disease Thiamine‐Responsive Megaloblastic Anemia (TRMA, OMIM #249270). Methods In vitro stimulated blood lymphocytes were used for cytogenetics and the isolation of genomic DNA used to perform whole exome sequencing (WES). To validate identified mutations, direct Sanger sequencing was performed following PCR amplification. Results A 6‐year‐old male born from a consanguineous couple presenting bone marrow failure and microcephaly was referred to our clinic for disease diagnosis. The patient presented a normal karyotype and no chromosomal fragility in response to DNA damage. WES analysis led to the identification of a new pathogenic variant in the SLC19A2 gene (c.596C>G, pSer199Ter) allowing to identify the young boy as a TRMA patient. Conclusion Our analysis extend the number of inactivating mutations in SLC19A2 leading to TRMA that could guide future prenatal diagnosis for the family and follow‐up for patients.

Published
2019

41. Familial predisposition to TP53/complex karyotype MDS and leukemia in DNA repair-deficient xeroderma pigmentosum

Author

Nathalie Droin, Alain Sarasin, Jean-Luc Schmutz, Stéphane de Botton, Samuel Quentin, Anna Raimbault, Filippo Rosselli, Alain Taieb, Véronique Saada, Vahid Asnafi, Jean Soulier, Yannick Boursin, Philippe Dessen, Patricia Kannouche, Thierry Leblanc, Nathalie Auger, Caroline Robert, Flore Sicre de Fontbrune, Mourad Sahbatou, Laurianne Drieu La Rochelle, Marie Sebert, Carlos Frederico Martins Menck, Eric Solary, Génomes et cancer (GC (FRE2939)), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Genetique et Biotherapies des Maladies Degeneratives et Proliferatives du Systeme Nerveux (Inserm U745), Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Bourgogne (UB), Fondation Jean Dausset CEPH, Laboratory of Hematology, Gustave Roussy, Villejuif, Praxiling (Praxiling), Centre National de la Recherche Scientifique (CNRS)-Université Paul-Valéry - Montpellier 3 (UPVM), Plateforme de Bioinformatique [Gustave Roussy], Analyse moléculaire, modélisation et imagerie de la maladie cancéreuse (AMMICa), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Hématopoïèse normale et pathologique (U1170 Inserm), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut Necker Enfants-Malades (INEM - UM 111 (UMR 8253 / U1151)), Service de Dermatologie et Allergologie [CHRU Nancy], Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy), Service de génétique médicale, Université de Bordeaux (UB)-CHU Bordeaux [Bordeaux]-Groupe hospitalier Pellegrin, Universidade de Sao Paulo, Institute of Biomedical Sciences, Universidade de São Paulo (USP)-Institute of Biomedical Sciences (ICB/USP), Universidade de São Paulo (USP), Stabilité Génétique et Oncogenèse (UMR 8200), Hematopoïèse et Cellules Souches (U362), Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Hôpital Saint-Louis, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Diderot - Paris 7 (UPD7), Service d'hématologie et immunologie pédiatrique, Université Paris Diderot - Paris 7 (UPD7)-Hôpital Robert Debré-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Institut Gustave Roussy (IGR), Radiothérapie moléculaire (UMR 1030), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Gustave Roussy (IGR)-Université Paris-Sud - Paris 11 (UP11), 3UMR728 INSERM Unité d'immuno-hématologie (UIH) and laboratoire d'hématologie, Hôpital St-Louis, AP-HP, Centre National de la Recherche Scientifique (CNRS), Unité d'Hémato-Immunologie pédiatrique [Hôpital Robert Debré, Paris], Service d'Immuno-hématologie pédiatrique [Hôpital Robert Debré, Paris], Hôpital Robert Debré-Hôpital Robert Debré, Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL (ENSCP)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL (ENSCP)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Praxiling UMR 5267 (Praxiling), Université Paul-Valéry - Montpellier 3 (UM3)-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Diderot - Paris 7 (UPD7)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP), and Université Paris Diderot - Paris 7 (UPD7)-Hôpital Robert Debré-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)

Subjects
0301 basic medicine, Xeroderma pigmentosum, DNA repair, [SDV]Life Sciences [q-bio], Immunology, medicine.disease_cause, Biochemistry, 03 medical and health sciences, 0302 clinical medicine, Complex Karyotype, Familial predisposition, medicine, Letter to Blood, Gene, ComputingMilieux_MISCELLANEOUS, Genetics, Mutation, business.industry, Cell Biology, Hematology, medicine.disease, 3. Good health, Leukemia, 030104 developmental biology, 030220 oncology & carcinogenesis, business, Founder effect
Abstract

There is a Blood Commentary on this article in this issue.

Published
2019

42. Light-induced formation of NO in endothelial cells by photoactivatable NADPH analogues targeting nitric-oxide synthase

Author

Nhi Ha Nguyen, Patrick Tauc, Jean-Luc Boucher, Françoise Simon, Rahima Chennoufi, Bertrand Cinquin, Juan Xie, Nicolas Bogliotti, Aimeric Cabrié, Eric Deprez, Anny Slama-schwok, Laboratoire de Biologie et de Pharmacologie Appliquée (LBPA), École normale supérieure - Cachan (ENS Cachan)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Photophysique et Photochimie Supramoléculaires et Macromoléculaires (PPSM), École normale supérieure - Cachan (ENS Cachan)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Biotechnologie et Pharmacogénétique Appliquée (LBPA), Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques (LCBPT - UMR 8601), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), CCSD, Accord Elsevier, and Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité)

Subjects
0301 basic medicine, Cell death, Photoactivation, Nitric-oxide synthase activator, Light, Nitric Oxide Synthase Type III, Endothelial cells, Biophysics, Golgi Apparatus, [SDV.BBM.BP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, Nitric Oxide, Biochemistry, 03 medical and health sciences, symbols.namesake, chemistry.chemical_compound, 0302 clinical medicine, Enos, Ribose, Human Umbilical Vein Endothelial Cells, medicine, Humans, Viability assay, Molecular Biology, Nicotinamide, biology, Two-photon absorption probes, Golgi apparatus, biology.organism_classification, Adenosine, Nitric oxide synthase, [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, 030104 developmental biology, chemistry, 030220 oncology & carcinogenesis, symbols, biology.protein, Photo-induced electron transfer, NADP, Intracellular, medicine.drug
Abstract

Background Nitric-oxide synthases (NOS) catalyze the formation of NO using NADPH as electron donor. We have recently designed and synthesized a new series of two-photon absorbing and photoactivatable NADPH analogues (NT). These compounds bear one or two carboxymethyl group(s) on the 2′- or/and 3′-position(s) of the ribose in the adenosine moiety, instead of a 2′-phosphate group, and differ by the nature of the electron donor in their photoactivatable chromophore (replacing the nicotinamide moiety). Here, we addressed the ability of NTs to photoinduce eNOS-dependent NO production in endothelial cells. Methods The cellular fate of NTs and their photoinduced effects were studied using multiphoton fluorescence imaging, cell viability assays and a BODIPY-derived NO probe for NO measurements. The eNOS dependence of photoinduced NO production was addressed using two NOS inhibitors (NS1 and L-NAME) targeting the reductase and the oxygenase domains, respectively. Results We found that, two compounds, those bearing a single carboxymethyl group on the 3′-position of the ribose, colocalize with the Golgi apparatus (the main intracellular location of eNOS) and display high intracellular two-photon brightness. Furthermore, a eNOS-dependent photooxidation was observed for these two compounds only, which is accompanied by a substantial intracellular NO production accounting for specific photocytotoxic effects. Conclusions We show for the first time that NT photoactivation efficiently triggers electron flow at the eNOS level and increases the basal production of NO by endothelial cells. General significance Efficient photoactivatable NADPH analogues targeting NOS could have important implications for generating apoptosis in tumor cells or modulating NO-dependent physiological processes.

Published
2019

43. A journey with common fragile sites: From S phase to telophase

Author

Filippo Rosselli, Michelle Debatisse, Dynamique de l'information génétique : bases fondamentales et cancer (DIG CANCER), Centre National de la Recherche Scientifique (CNRS)-Institut Curie [Paris]-Université Pierre et Marie Curie - Paris 6 (UPMC), Stabilité Génétique et Oncogenèse (UMR 8200), and Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Genome instability, Cancer Research, Chromosomal fragile site, Chromosome Fragile Sites, [SDV]Life Sciences [q-bio], Biology, Genome, FANC proteins, S Phase, Evolution, Molecular, 03 medical and health sciences, 0302 clinical medicine, Transcription (biology), Evolutionary biology, 030220 oncology & carcinogenesis, Genetics, Animals, Humans, Telophase, Mitosis, Gene, ComputingMilieux_MISCELLANEOUS
Abstract

Some regions of the genome, notably common fragile sites (CFSs), are hypersensitive to replication stress and often involved in the generation of gross chromosome rearrangements in cancer cells. CFSs nest within very large genes and display cell-type-dependent instability. Fragile or not, large genes tend to replicate late in S-phase. A number of data now show that transcription perturbs replication completion across the body of large genes, particularly upon replication stress. However, the molecular mechanisms by which transcription elicits such under-replication and subsequent instability remain unclear. We present here our view of the mechanisms responsible for CFS under-replication and those allowing the cells to cope with this problem in G2 and mitosis. We notably focus on the major role played by the FANC proteins in the protection of CFSs from S phase up to late mitosis. We finally discuss a possible rationale for the conservation of large genes across vertebrate evolution.

Published
2019

44. SMC5/6 acts jointly with Fanconi anemia factors to support DNA repair and genome stability

Author

Francesco Rossi, Dongyi Xu, Xinlin Xu, Pierre Devulder, Nanda Kumar Jegadesan, Minoru Takata, Filippo Rosselli, Takuya Abe, Dana Branzei, Ryotaro Kawasumi, Anne Helbling-Leclerc, Intégrité du génome et cancers (IGC), Institut Gustave Roussy (IGR)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Laboratoire des Sciences de l'Information et des Systèmes (LSIS), Centre National de la Recherche Scientifique (CNRS)-Arts et Métiers Paristech ENSAM Aix-en-Provence-Université de Toulon (UTLN)-Aix Marseille Université (AMU), Tohoku University [Sendai], State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, PeKing University, Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Aix Marseille Université (AMU)-Université de Toulon (UTLN)-Arts et Métiers Paristech ENSAM Aix-en-Provence-Centre National de la Recherche Scientifique (CNRS), and Peking University [Beijing]

Subjects
congenital, hereditary, and neonatal diseases and abnormalities, DNA Repair, DNA repair, Chromosomal Proteins, Non-Histone, [SDV]Life Sciences [q-bio], RAD51, Cell Cycle Proteins, SMC5/6, Biochemistry, Genomic Instability, Article, DEAD-box RNA Helicases, 03 medical and health sciences, 0302 clinical medicine, Fanconi anemia, hemic and lymphatic diseases, FANCD2, Genetics, medicine, Humans, Molecular Biology of Disease, FANCM, Molecular Biology, Mitosis, Gene, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, intra‐ and inter‐strand crosslinks, mitotic instability, biology, Cell Cycle, DNA Helicases, Helicase, nutritional and metabolic diseases, DNA Replication, Repair & Recombination, Articles, medicine.disease, Cell biology, Fanconi Anemia, biology.protein, 030217 neurology & neurosurgery, DNA Damage, HeLa Cells
Abstract

SMC5/6 function in genome integrity remains elusive. Here, we show that SMC5 dysfunction in avian DT40 B cells causes mitotic delay and hypersensitivity toward DNA intra‐ and inter‐strand crosslinkers (ICLs), with smc5 mutants being epistatic to FANCC and FANCM mutations affecting the Fanconi anemia (FA) pathway. Mutations in the checkpoint clamp loader RAD17 and the DNA helicase DDX11, acting in an FA‐like pathway, do not aggravate the damage sensitivity caused by SMC5 dysfunction in DT40 cells. SMC5/6 knockdown in HeLa cells causes MMC sensitivity, increases nuclear bridges, micronuclei, and mitotic catastrophes in a manner similar and non‐additive to FANCD2 knockdown. In both DT40 and HeLa systems, SMC5/6 deficiency does not affect FANCD2 ubiquitylation and, unlike FANCD2 depletion, RAD51 focus formation. SMC5/6 components further physically interact with FANCD2‐I in human cells. Altogether, our data suggest that SMC5/6 functions jointly with the FA pathway to support genome integrity and DNA repair and may be implicated in FA or FA‐related human disorders., The structural maintenance complex SMC5/6 interacts with and functions jointly with key components of the Fanconi anemia pathway to facilitate DNA repair and preserve genome stability.

Published
2019

45. Why is immunotherapy effective (or not) in patients with MSI/MMRD tumors?

Author

Laetitia Nebot-Bral, Patricia Kannouche, Clelia Coutzac, Nathalie Chaput, Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d’Immunomonitoring en Oncologie (LIO), Analyse moléculaire, modélisation et imagerie de la maladie cancéreuse (AMMICa), Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and Kannouche, patricia

Subjects
0301 basic medicine, Cancer Research, medicine.medical_treatment, Programmed Cell Death 1 Receptor, Context (language use), [SDV.CAN]Life Sciences [q-bio]/Cancer, [SDV.GEN] Life Sciences [q-bio]/Genetics, DNA Mismatch Repair, B7-H1 Antigen, Mutation Accumulation, 03 medical and health sciences, 0302 clinical medicine, Immune system, [SDV.CAN] Life Sciences [q-bio]/Cancer, Antigen, Neoplasms, Tumor Microenvironment, medicine, Humans, CTLA-4 Antigen, Radiology, Nuclear Medicine and imaging, ComputingMilieux_MISCELLANEOUS, Tumor microenvironment, [SDV.GEN]Life Sciences [q-bio]/Genetics, [SDV.MHEP] Life Sciences [q-bio]/Human health and pathology, business.industry, Microsatellite instability, Hematology, General Medicine, Immunotherapy, Prognosis, medicine.disease, Immune checkpoint, 3. Good health, 030104 developmental biology, Oncology, 030220 oncology & carcinogenesis, Cancer research, Microsatellite Instability, DNA mismatch repair, business, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract

In the last few years, immunotherapy has revolutionized the oncology landscape by targeting the host immune system. Blocking immune checkpoints such as cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4), programmed cell death-1 (PD-1) and its ligand (PD-L1 or B7-H1), has proven its efficacy in several solid cancers. Recently, several clinical studies have demonstrated a significant improvement in clinical response to the anti-PD-1-based immunotherapy in a subset of patients with microsatellite instability-high (MSI-H)/mismatch repair (MMR)-deficient tumors that accumulate short insertion/deletion mutations notably in coding microsatellites regions of the genome. Thus, the responsiveness of MSI cancers to immune checkpoint inhibitors can be explained by the increased rate of putative frameshift peptide neoantigens and the immunogenic tumor microenvironment. However, not all MSI tumors respond to immunotherapy. The current review will summarize how and why MMR deficiency has emerged as an important predictor of sensitivity for immunotherapy-based strategies. We will also discuss tumor-cell intrinsic genetic and immune-related features of MSI tumors that can modulate immune checkpoint blockade response and explain primary and/or acquired resistance to anti-PD-1 therapy. Finally, we will also discuss about emerging scores which can define more precisely the immune context of the tumor microenvironment and thus better evaluate prognosis and predict response to Immune Checkpoint Blockade.

Published
2019

46. Oxidative stress in thyroid carcinomas: biological and clinical significance

Author

Corinne Dupuy, Sophie Leboulleux, Camille Buffet, Rabii Ameziane El Hassani, Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut Gustave Roussy (IGR), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS), and Département d'imagerie médicale [Gustave Roussy]

Subjects
Genome instability, Cancer Research, DNA damage, Endocrinology, Diabetes and Metabolism, [SDV]Life Sciences [q-bio], Thyroid Gland, [SDV.CAN]Life Sciences [q-bio]/Cancer, Biology, medicine.disease_cause, Genomic Instability, Thyroid carcinoma, 03 medical and health sciences, 0302 clinical medicine, Endocrinology, Chromosome instability, medicine, Animals, Humans, Thyroid Neoplasms, Thyroid cancer, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, Thyroid, NADPH Oxidases, Oncogenes, Cell Dedifferentiation, medicine.disease, 3. Good health, Gene Expression Regulation, Neoplastic, Oxidative Stress, medicine.anatomical_structure, Oncology, 030220 oncology & carcinogenesis, Cancer research, Carcinogenesis, Reactive Oxygen Species, Oxidative stress, DNA Damage
Abstract

At physiological concentrations, reactive oxygen species (ROS), including superoxide anions and H2O2, are considered as second messengers that play key roles in cellular functions, such as proliferation, gene expression, host defence and hormone synthesis. However, when they are at supraphysiological levels, ROS are considered potent DNA-damaging agents. Their increase induces oxidative stress, which can initiate and maintain genomic instability. The thyroid gland represents a good model for studying the impact of oxidative stress on genomic instability. Indeed, one particularity of this organ is that follicular thyroid cells synthesise thyroid hormones through a complex mechanism that requires H2O2. Because of their detection in thyroid adenomas and in early cell transformation, both oxidative stress and DNA damage are believed to be neoplasia-preceding events in thyroid cells. Oxidative DNA damage is, in addition, detected in the advanced stages of thyroid cancer, suggesting that oxidative lesions of DNA also contribute to the maintenance of genomic instability during the subsequent phases of tumourigenesis. Finally, ionizing radiation and the mutation of oncogenes, such as RAS and BRAF, play a key role in thyroid carcinogenesis through separate and unique mechanisms: they upregulate the expression of two distinct ‘professional’ ROS-generating systems, the NADPH oxidases DUOX1 and NOX4, which cause DNA damage that may promote chromosomal instability, tumourigenesis and dedifferentiation.

Published
2019

47. Combining Homologous Recombination and Phosphopeptide-binding Data to Predict the Impact of BRCA1 BRCT Variants on Cancer Risk

Author

Petitalot, Ambre, Dardillac, Elodie, Jacquet, Eric, Nhiri, Naima, Guirouilh-Barbat, Josee, Julien, Patrick, Bouazzaoui, Isslam, Bonte, Dorine, Feunteun, Jean, Schnell, Jeff A., Lafitte, Philippe, Aude, Jean-Christophe, Nogues, Catherine, Rouleau, Etienne, Lidereau, Rosette, Lopez, Bernard S., Zinn-Justin, Sophie, Caputo, Sandrine M., Bonnet, Francoise, Jones, Natalie, Bubien, Virginie, Longy, Michel, Sevenet, Nicolas, Krieger, Sophie, Casters, Laurent, Vaur, Dominique, Uhrhammer, Nancy, Bignon, Yves Jean, Lizard, Sarab, Dumont, Aurelie, Revillion, Francoise, Leone, Melanie, Boutry-Kryza, Nadia, Sinilnikova, Olga, Remenieras, Audrey, Bourdon, Violaine, Noguchi, Tetsuro, Sobol, Hagay, Harmand, Pierre-Olivier, Vilquin, Paul, Pujol, Pascal, Jonveaux, Philippe, Bronner, Myriam, Sokolowska, Joanna, Delnatte, Capucine, Guibert, Virginie, Garrec, Celine, Bezieau, Stephan, Soubrier, Florent, Guillerm, Erell, Coulet, Florence, Lefol, Cedrick, Caux-Moncoutier, Virginie, Golmard, Lisa, Houdayer, Claude, Stoppa-Lyonnet, Dominique, Delvincoun, Chantal, Beaudoux, Olivia, Muller, Daniele, Toulas, Christine, Guillaud-Bataille, Marine, Bressac-De Paillerets, Brigitte, Unité de génétique et biologie des cancers (U830), Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11), Ligue Nationale Contre le Cancer - Paris, Ligue Nationnale Contre le Cancer, Institut de Chimie des Substances Naturelles (ICSN), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), BioInformatique Moléculaire (BIM), Département Biologie des Génomes (DBG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Enveloppe Nucléaire, Télomères et Réparation de l’ADN (INTGEN), Département Biochimie, Biophysique et Biologie Structurale (B3S), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie-Institut National de la Santé et de la Recherche Médicale (INSERM), Institute of Cellular and Molecular Radiobiology, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Centre National de la Recherche Scientifique (CNRS), Hématopoïèse normale et pathologique, Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Gustave Roussy (IGR)-Université Paris-Sud - Paris 11 (UP11), Laboratoire de Biologie Intégrative (LBI), Institut Curie, CRLCC René Huguenin, Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay, Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay-Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay-Institut de Biologie Intégrative de la Cellule (I2BC), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay-Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay, DIEP/DSV (DIEP/DSV), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Curie [Paris], Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Ligue Nationale Contre le Cancer (LNCC), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), and ANR-16-CE12-0011,2R-POL,ADN polymérase theta : lien entre réplication et réparation de l'ADN(2016)

Subjects
0301 basic medicine, Cancer Research, DNA repair, functional-analysis, [SDV]Life Sciences [q-bio], domain, Computational biology, Biology, medicine.disease_cause, 03 medical and health sciences, 0302 clinical medicine, medicine, ovarian, Molecular Biology, Cellular localization, Mutation, dna-damage, Phosphopeptide, Cancer, hereditary breast, structural basis, medicine.disease, 3. Good health, 030104 developmental biology, BRCT domain, Oncology, classification, 030220 oncology & carcinogenesis, repair, missense variants, Cancer research, Homologous recombination, Carcinogenesis, bach1 phosphopeptide
Abstract

BRCA1 mutations have been identified that increase the risk of developing hereditary breast and ovarian cancers. Genetic screening is now offered to patients with a family history of cancer, to adapt their treatment and the management of their relatives. However, a large number of BRCA1 variants of uncertain significance (VUS) are detected. To better understand the significance of these variants, a high-throughput structural and functional analysis was performed on a large set of BRCA1 VUS. Information on both cellular localization and homology-directed DNA repair (HR) capacity was obtained for 78 BRCT missense variants in the UMD-BRCA1 database and measurement of the structural stability and phosphopeptide-binding capacities was performed for 42 mutated BRCT domains. This extensive and systematic analysis revealed that most characterized causal variants affect BRCT-domain solubility in bacteria and all impair BRCA1 HR activity in cells. Furthermore, binding to a set of 5 different phosphopeptides was tested: all causal variants showed phosphopeptide-binding defects and no neutral variant showed such defects. A classification is presented on the basis of mutated BRCT domain solubility, phosphopeptide-binding properties, and VUS HR capacity. These data suggest that HR-defective variants, which present, in addition, BRCT domains either insoluble in bacteria or defective for phosphopeptide binding, lead to an increased cancer risk. Furthermore, the data suggest that variants with a WT HR activity and whose BRCT domains bind with a WT affinity to the 5 phosphopeptides are neutral. The case of variants with WT HR activity and defective phosphopeptide binding should be further characterized, as this last functional defect might be sufficient per se to lead to tumorigenesis. Implications: The analysis of the current study on BRCA1 structural and functional defects on cancer risk and classification presented may improve clinical interpretation and therapeutic selection.

Published
2019

48. Selective modification of a native protein in a patient tissue homogenate using palladium nanoparticles

Author

Ruy A. N. Louzada, Benoit Lambert, Patrick Couvreur, Stéphanie Yen-Nicolaÿ, Didier Desmaële, Anaëlle Dumas, Arnaud Peramo, Shannon Pecnard, Jacques Young, Corinne Dupuy, Hynd Remita, Raphael Corre, Mireille Benoît, Institut Galien Paris-Saclay (IGPS), Institut de Chimie du CNRS (INC)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Institut Galien Paris-Sud (IGPS), Université Paris-Sud - Paris 11 (UP11)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie Physique D'Orsay (LCPO), Faculté de Pharmacie, IPSIT, Université Paris Sud, Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Institut Gustave Roussy (IGR), Laboratoire de l'intégration, du matériau au système (IMS), Université Sciences et Technologies - Bordeaux 1-Institut Polytechnique de Bordeaux-Centre National de la Recherche Scientifique (CNRS), U693, Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Sud (Paris 11), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), UMS IPSIT, Plate forme d'imagerie cellulaire, Université Paris-Saclay, Récepteurs stéroïdiens : physiopathologie endocrinienne et métabolique, Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR93-Université Paris-Sud - Paris 11 (UP11), and Physico-chimie, pharmacotechnie, biopharmacie (PCPB)

Subjects
[SDV]Life Sciences [q-bio], medicine.medical_treatment, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, 010402 general chemistry, 01 natural sciences, Catalysis, Materials Chemistry, medicine, Native protein, [CHIM]Chemical Sciences, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, ComputingMilieux_MISCELLANEOUS, Tissue homogenate, 010405 organic chemistry, Chemistry, Thyroid, Metals and Alloys, Palladium nanoparticles, [CHIM.CATA]Chemical Sciences/Catalysis, [CHIM.MATE]Chemical Sciences/Material chemistry, General Chemistry, 0104 chemical sciences, Surfaces, Coatings and Films, Electronic, Optical and Magnetic Materials, medicine.anatomical_structure, Biochemistry, Ceramics and Composites, Thyroglobulin
Abstract

International audience; We have developed new benign palladium nanoparticles able to catalyze the Suzuki-Miyaura cross-coupling reaction on human thyroglobulin (Tg), a naturally iodinated protein produced by the thyroid gland, in homogenates from patients' tissues. This represents the first example of a chemoselective native protein modification using transition metal nanoobjects in near-organ medium.

Published
2019

49. Slx5-Slx8 ubiquitin ligase targets active pools of the Yen1 nuclease to limit crossover formation

Author

Ibtissam Talhaoui, Manuel Bernal, Janet R. Mullen, Hugo Dorison, Benoit Palancade, Steven J. Brill, Gerard Mazón, Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Institut Jacques Monod (IJM (UMR_7592)), and Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Cell Nucleus, Saccharomyces cerevisiae Proteins, Lysine, Science, [SDV]Life Sciences [q-bio], education, Ubiquitination, Sumoylation, Saccharomyces cerevisiae, Article, Substrate Specificity, Chromosome Segregation, lcsh:Q, Crossing Over, Genetic, lcsh:Science, Cell Nucleolus, Gene Deletion, ComputingMilieux_MISCELLANEOUS, DNA Damage, Protein Binding
Abstract

The repair of double-stranded DNA breaks (DSBs) by homologous recombination involves the formation of branched intermediates that can lead to crossovers following nucleolytic resolution. The nucleases Mus81-Mms4 and Yen1 are tightly controlled during the cell cycle to limit the extent of crossover formation and preserve genome integrity. Here we show that Yen1 is further regulated by sumoylation and ubiquitination. In vivo, Yen1 becomes sumoylated under conditions of DNA damage by the redundant activities of Siz1 and Siz2 SUMO ligases. Yen1 is also a substrate of the Slx5-Slx8 ubiquitin ligase. Loss of Slx5-Slx8 stabilizes the sumoylated fraction, attenuates Yen1 degradation at the G1/S transition, and results in persistent localization of Yen1 in nuclear foci. Slx5-Slx8-dependent ubiquitination of Yen1 occurs mainly at K714 and mutation of this lysine increases crossover formation during DSB repair and suppresses chromosome segregation defects in a mus81∆ background., Nucleases are regulated during the cell cycle to control for crossover formation and maintain genome integrity. Here the authors reveal that the yeast Holliday junction resolvase Yen is a sumoylation target and it is regulated by the ubiquitin ligases Slx5/Slx8 during crossover formation.

Published
2018

50. TET2-mediated 5-hydroxymethylcytosine induces genetic instability and mutagenesis

Author

Lise Secardin, William Vainchenker, Olivier Bernard, Alexander A. Ishchenko, Murat Saparbaev, Emna Mahfoudhi, Philippe Rameau, Véronique Della Valle, Ibtissam Talhaoui, Isabelle Plo, Xenia Cabagnols, Salem Abbes, Hématopoïèse normale et pathologique (UMR 1170 ), Université Paris-Sud - Paris 11 (UP11) - Institut Gustave Roussy (IGR) - Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire d'hématologie moléculaire et cellulaire, institut Pasteur de Tunis, Institut Pasteur de Tunis - Réseau International des Instituts Pasteur, Institut Gustave Roussy (IGR), Réparation de l’ADN (CNRS UMR 8200), This work was supported by grants from Agence Nationale de la Recherche (ANR-Blanc 2013 GERMPN) to IP and [ANR Blanc 2010 Project ANR-10-BLAN-1617] to IP and MS, Association pour la Recherche contre le Cancer (ARC) (ARC libre 2012) to IP, Electricité de France (http://www.edf.fr) RB 2014-26 to MS and Fondation de France (http://www.fondationdefrance.org) [#2012 00029161] to AAI. Labex GR-Ex (IP, WV) is funded by the program 'Investissements d’avenir'. EM was funded by INSERM/DGRST then a grant from Institut Pasteur from Tunis, Tunisia. LS were supported by Ph.D grants from the Cancéropôle Région Ile de France (DIM cellule souche). XC was supported by Ph.D MENRT grant. IT was supported by postdoctoral fellowship from the Fondation ARC (http://www.arc-cancer.net) PDF20110603195., ANR-Blanc 2013 GERMPN, GERMPN, ANR-10-BLAN-1617, EPIGENOME, Formation, dynamics and epigenetic functions of 5-hydroxymethylcytosine residues in DNA.(2010), Hématopoïèse normale et pathologique (U1170 Inserm), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Tunis El Manar (UTM), Laboratoire d'hématologie moléculaire et cellulaire (LR11IPT07), Institut Pasteur de Tunis, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Plateforme imagerie et cytométrie (PFIC), Analyse moléculaire, modélisation et imagerie de la maladie cancéreuse (AMMICa), Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire d'Hématologie, Institut Pasteur de Tunis, This work was supported by grants from Agence Nationale de la Recherche (ANR-Blanc 2013 GERMPN) to IP and [ANR Blanc 2010 Project ANR-10-BLAN-1617] to IP and MS, Association pour la Recherche contre le Cancer (ARC) (ARC libre 2012) to IP, Electricité de France (http://www.edf.fr) RB 2014-26 to MS and Fondation de France (http://www.fondationdefrance.org) [#2012 00029161] to AAI. Labex GR-Ex (IP, WV) is funded by the program 'Investissements d’avenir'. EM was funded by INSERM/DGRST then a grant from Institut Pasteur from Tunis, Tunisia. LS were supported by Ph.D grants from the Cancéropôle Région Ile de France (DIM cellule souche). XC was supported by Ph.D MENRT grant. IT was supported by postdoctoral fellowship from the Fondation ARC., We thank the cytometry platform of Gustave Roussy (Y. Lecluse)., ANR-10-BLAN-1617,EPIGENOME,Formation, dynamics and epigenetic functions of 5-hydroxymethylcytosine residues in DNA.(2010), ANR-13-JSV1-0007,GERMPN,Etude des cas familiaux de néoplasmes myéloproliférifs: recherche des anomalies génétiques et de leurs fonctions.(2013), Pasteur Tunis, Institut, BLANC - Formation, dynamics and epigenetic functions of 5-hydroxymethylcytosine residues in DNA. - - EPIGENOME2010 - ANR-10-BLAN-1617 - BLANC - VALID, Jeunes Chercheuses et Jeunes Chercheurs - Etude des cas familiaux de néoplasmes myéloproliférifs: recherche des anomalies génétiques et de leurs fonctions. - - GERMPN2013 - ANR-13-JSV1-0007 - JC - VALID, and Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, DNA Repair, Genetic instability, MESH: Fibroblasts/cytology, MESH: Tumor Suppressor Protein p53/metabolism, MESH: Fibroblasts/metabolism, MESH: Mutagenesis, MESH: Base Sequence, MESH: Thymine DNA Glycosylase/deficiency, Biochemistry, MESH: DNA Repair, Epigenesis, Genetic, S Phase, MESH: Thymine DNA Glycosylase/genetics, Mice, chemistry.chemical_compound, MESH: Genomic Instability, MESH: DNA-Binding Proteins/genetics, MESH: B-Lymphocytes/metabolism, MESH: Animals, MESH: Cytosine/metabolism, MESH: Epigenesis, Genetic, MESH: Megakaryocyte Progenitor Cells/cytology, MESH: B-Lymphocytes/cytology, B-Lymphocytes, [SDV.MHEP] Life Sciences [q-bio]/Human health and pathology, MESH: Cytosine/analogs & derivatives, MESH: S Phase, Base excision repair, DNA-Binding Proteins, MESH: Proto-Oncogene Proteins/genetics, 5-hmC, CpG site, 5-Methylcytosine, [SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Cytosine, Guanine, MESH: Hydroxylation, TDG, Cell cycle, Biology, MESH: DNA-Binding Proteins/metabolism, Hydroxylation, Genomic Instability, Cell Line, Dioxygenases, 03 medical and health sciences, MESH: Tumor Suppressor Protein p53/genetics, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Proto-Oncogene Proteins, Animals, Humans, MESH: Mice, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, Molecular Biology, Megakaryocyte Progenitor Cells, MESH: 5-Methylcytosine/analogs & derivatives, 5-Hydroxymethylcytosine, TET2, MESH: Humans, Base Sequence, MESH: Proto-Oncogene Proteins/metabolism, Mutagenesis, MESH: 5-Methylcytosine/metabolism, Cell Biology, Fibroblasts, Molecular biology, Thymine DNA Glycosylase, MESH: Cell Line, 030104 developmental biology, DNA demethylation, chemistry, DNA glycosylase, MESH: Megakaryocyte Progenitor Cells/metabolism, Tumor Suppressor Protein p53
Abstract

International audience; The family of Ten-Eleven Translocation (TET) proteins is implicated in the process of active DNA demethy-lation and thus in epigenetic regulation. TET 1, 2 and 3 proteins are oxygenases that can hydroxylate 5-methylcytosine (5-mC) into 5-hydroxymethylcytosine (5-hmC) and further oxidize 5-hmC into 5-formylcytosine (5-fC) and 5-carboxylcytosine (5-caC). The base excision repair (BER) pathway removes the resulting 5-fC and 5-caC bases paired with a guanine and replaces them with regular cytosine. The question arises whether active modification of 5-mC residues and their subsequent elimination could affect the genomic DNA stability. Here, we generated two inducible cell lines (Ba/F3-EPOR, and UT7) over-expressing wild-type or catalytically inactive human TET2 proteins. Wild-type TET2 induction resulted in an increased level of 5-hmC and a cell cycle defect in S phase associated with higher level of phospho-rylated P53, chromosomal and centrosomal abnormalities. Furthermore, in a thymine-DNA glycosylase (Tdg) deficient context, the TET2-mediated increase of 5-hmC induces mutagenesis characterized by GC > AT transitions in CpG context suggesting a mutagenic potential of 5-hmC metabolites. Altogether, these data suggest that TET2 activity and the levels of 5-hmC and its derivatives should be tightly controlled to avoid genetic and chromosomal instabilities. Moreover, TET2-mediated active demethylation might be a very dangerous process if used to entirely demethylate the genome and might rather be used only at specific loci.

Published
2016

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