37 results on '"Juan, Gerez"'
Search Results
2. Non-invasive imaging of tau-targeted probe uptake by whole brain multi-spectral optoacoustic tomography
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Roland Riek, Artur Luzgin, Uwe Konietzko, Juan Gerez, Daniel Razansky, Zhenyue Chen, Maiko Ono, Bin Ji, Xosé Luís Deán-Ben, Patrick Vagenknecht, Roger M. Nitsch, Cinzia A Maschio, Daniela Noain, Jens Sobek, Makoto Higuchi, Jan Klohs, Ruiqing Ni, University of Zurich, Dean-Ben, Xose Luis, and Ni, Ruiqing
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Pathology ,medicine.medical_specialty ,Animal model ,Fluorescence imaging ,Frontotemporal dementia ,Optoacoustic imaging ,Tau ,Mice, Transgenic ,tau Proteins ,610 Medicine & health ,Progressive supranuclear palsy ,170 Ethics ,Mice ,chemistry.chemical_compound ,Neuroimaging ,Alzheimer Disease ,In vivo ,medicine ,Animals ,Humans ,Corticobasal degeneration ,2741 Radiology, Nuclear Medicine and Imaging ,Radiology, Nuclear Medicine and imaging ,10237 Institute of Biomedical Engineering ,Brain ,General Medicine ,Human brain ,11359 Institute for Regenerative Medicine (IREM) ,medicine.disease ,Disease Models, Animal ,medicine.anatomical_structure ,Tauopathies ,chemistry ,Positron-Emission Tomography ,Thioflavin ,Tauopathy ,Alzheimer's disease - Abstract
Purpose Abnormal tau accumulation within the brain plays an important role in tauopathies such as Alzheimer’s disease and frontotemporal dementia. High-resolution imaging of tau deposits at the whole-brain scale in animal disease models is highly desired. Methods We approached this challenge by non-invasively imaging the brains of P301L mice of 4-repeat tau with concurrent volumetric multi-spectral optoacoustic tomography (vMSOT) at ~ 115 μm spatial resolution using the tau-targeted pyridinyl-butadienyl-benzothiazole derivative PBB5 (i.v.). In vitro probe characterization, concurrent vMSOT and epi-fluorescence imaging of in vivo PBB5 targeting (i.v.) was performed in P301L and wild-type mice, followed by ex vivo validation using AT-8 antibody for phosphorylated tau. Results PBB5 showed specific binding to recombinant K18 tau fibrils by fluorescence assay, to post-mortem Alzheimer’s disease brain tissue homogenate by competitive binding against [11C]PBB3 and to tau deposits (AT-8 positive) in post-mortem corticobasal degeneration and progressive supranuclear palsy brains. Dose-dependent optoacoustic and fluorescence signal intensities were observed in the mouse brains following i.v. administration of different concentrations of PBB5. In vivo vMSOT brain imaging of P301L mice showed higher retention of PBB5 in the tau-laden cortex and hippocampus compared to wild-type mice, as confirmed by ex vivo vMSOT, epi-fluorescence, multiphoton microscopy, and immunofluorescence staining. Conclusions We demonstrated non-invasive whole-brain imaging of tau in P301L mice with vMSOT system using PBB5 at a previously unachieved ~ 115 μm spatial resolution. This platform provides a new tool to study tau spreading and clearance in a tauopathy mouse model, foreseeable in monitoring tau targeting putative therapeutics., European Journal of Nuclear Medicine and Molecular Imaging, 49 (7), ISSN:1619-7070, ISSN:1619-7089
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- 2022
3. Co-Expression of Wild-Type P2X7R with Gln460Arg Variant Alters Receptor Function.
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Fernando Aprile-Garcia, Michael W Metzger, Marcelo Paez-Pereda, Herbert Stadler, Matías Acuña, Ana C Liberman, Sergio A Senin, Juan Gerez, Esteban Hoijman, Damian Refojo, Mišo Mitkovski, Markus Panhuysen, Walter Stühmer, Florian Holsboer, Jan M Deussing, and Eduardo Arzt
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Medicine ,Science - Abstract
The P2X7 receptor is a member of the P2X family of ligand-gated ion channels. A single-nucleotide polymorphism leading to a glutamine (Gln) by arginine (Arg) substitution at codon 460 of the purinergic P2X7 receptor (P2X7R) has been associated with mood disorders. No change in function (loss or gain) has been described for this SNP so far. Here we show that although the P2X7R-Gln460Arg variant per se is not compromised in its function, co-expression of wild-type P2X7R with P2X7R-Gln460Arg impairs receptor function with respect to calcium influx, channel currents and intracellular signaling in vitro. Moreover, co-immunoprecipitation and FRET studies show that the P2X7R-Gln460Arg variant physically interacts with P2X7R-WT. Specific silencing of either the normal or polymorphic variant rescues the heterozygous loss of function phenotype and restores normal function. The described loss of function due to co-expression, unique for mutations in the P2RX7 gene so far, explains the mechanism by which the P2X7R-Gln460Arg variant affects the normal function of the channel and may represent a mechanism of action for other mutations.
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- 2016
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4. α-Synuclein aggregation nucleates through liquid–liquid phase separation
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Sandhya Bhatia, Amrendra K. Singh, Jaladhar Mahato, Soumik Ray, Ranjith Padinhateeri, Samir K. Maji, Laxmikant G. Gadhe, Siddhartha Maiti, Ambuja Navalkar, Roland Riek, Debdeep Chatterjee, Rajlaxmi Panigrahi, G. Krishnamoorthy, Nitu Singh, Sushil Kumar Pandey, Rakesh Kumar, Arindrajit Chowdhury, Komal Patel, Surabhi Mehra, Juan Gerez, Debalina Datta, and Ashutosh Kumar
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Amyloid ,010405 organic chemistry ,Chemistry ,animal diseases ,General Chemical Engineering ,Mutagenesis ,General Chemistry ,010402 general chemistry ,01 natural sciences ,In vitro ,nervous system diseases ,0104 chemical sciences ,Aggresome ,nervous system ,Microtubule ,Biophysics ,Liquid liquid ,heterocyclic compounds ,α synuclein ,Cellular model - Abstract
α-Synuclein (α-Syn) aggregation and amyloid formation is directly linked with Parkinson's disease pathogenesis. However, the early events involved in this process remain unclear. Here, using the in vitro reconstitution and cellular model, we show that liquid-liquid phase separation of α-Syn precedes its aggregation. In particular, in vitro generated α-Syn liquid-like droplets eventually undergo a liquid-to-solid transition and form an amyloid hydrogel that contains oligomers and fibrillar species. Factors known to aggravate α-Syn aggregation, such as low pH, phosphomimetic substitution and familial Parkinson's disease mutations, also promote α-Syn liquid-liquid phase separation and its subsequent maturation. We further demonstrate α-Syn liquid-droplet formation in cells. These cellular α-Syn droplets eventually transform into perinuclear aggresomes, the process regulated by microtubules. This work provides detailed insights into the phase-separation behaviour of natively unstructured α-Syn and its conversion to a disease-associated aggregated state, which is highly relevant in Parkinson's disease pathogenesis.
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- 2020
5. Non-invasive optoacoustic imaging of tau in P301L mice
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Juan Gerez, Xosé Luís Deán-Ben, Patrick Vagenknecht, Zhenyue Chen, Daniel Razansky, Roger M. Nitsch, Jan Klohs, Ruiqing Ni, and University of Zurich
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Materials science ,medicine.diagnostic_test ,Non invasive ,10050 Institute of Pharmacology and Toxicology ,610 Medicine & health ,medicine.disease ,170 Ethics ,Three dimensional imaging ,Optical imaging ,Nuclear magnetic resonance ,Positron emission tomography ,mental disorders ,medicine ,10237 Institute of Biomedical Engineering ,Surface plasmon resonance ,Optoacoustic imaging ,Preclinical imaging ,Frontotemporal dementia - Abstract
Abnormal tau accumulation plays an important role in Alzheimer’s disease and frontotemporal dementia. There is a gap in preclinical high-resolution imaging of tau. Here we showed non-invasive whole-brain optoacoustic tau imaging in P301L tau mice.
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- 2021
6. Transcranial detection of tauopathy in vivo in P301L mice with high‐resolution large‐field multifocal illumination fluorescence microscopy
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Gloria Shi, Roland Riek, K. Peter R. Nilsson, Daniel Razansky, Zhenyue Chen, Jan Klohs, Ruiqing Ni, Quanyu Zhou, and Juan Gerez
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Fluorescence-lifetime imaging microscopy ,Epidemiology ,Health Policy ,medicine.disease ,In vitro ,Psychiatry and Mental health ,Cellular and Molecular Neuroscience ,chemistry.chemical_compound ,Developmental Neuroscience ,chemistry ,In vivo ,medicine ,Fluorescence microscope ,Biophysics ,Immunohistochemistry ,Thioflavin ,Neurology (clinical) ,Tauopathy ,Geriatrics and Gerontology ,Intravital microscopy - Abstract
Background Tau abnormal aggregates have been visualized in‐vivo using intravital microscopy techniques with high resolution, but are limited to a small field‐of‐view and depth. Here, we report a new method for the transcranial detection of tau deposits accross the brain with 6 μm resolution. Method In vitro Thioflavin T fluorescence assay was performed to screen fluorescence imaging probes that bind to tau fibrils. P301L (Thy1.2) mouse models of frontal temporal lobe dementia with four repeat tau and non‐transgenic littermates were imaged in vivo using a novel large‐field multifocal illumination (LMI) fluorescence microscopy technique using luminescent conjugated oligothiophenes h‐FTAA. Immunohistochemical staining were performed on P301L mouse brain tissue sections using anti‐phosphorylated tau antibodies AT8 and AT100. Result In vitro Thioflavin T fluorescence assay and LMI imaging shows that h‐FTAA bind to recombinant full‐length tau fibrils. In vivo LMI imaging using h‐FTAA showed higher retention in the cortex of P301L mice at 10 month‐of‐age compared to age‐matched non‐transgenic littermates. Immunohistochemical staining on P301L mouse brain tissue sections showed co‐localization of h‐FTAA with anti‐phosphorylated tau antibodies AT8 and AT100, thus verifying the specificity of the probe to tauopathy in P301L mice. Conclusion We demonstrate a new in vivo high‐resolution imaging platform for detection of tauopathy in murine models of frontal temporal lobe tauopathy.
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- 2020
7. In silico structural and functional characterization of the RSUME splice variants.
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Juan Gerez, Mariana Fuertes, Lucas Tedesco, Susana Silberstein, Gustavo Sevlever, Marcelo Paez-Pereda, Florian Holsboer, Adrián G Turjanski, and Eduardo Arzt
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Medicine ,Science - Abstract
RSUME (RWD-containing SUMO Enhancer) is a small protein that increases SUMO conjugation to proteins. To date, four splice variants that codify three RSUME isoforms have been described, which differ in their C-terminal end. Comparing the structure of the RSUME isoforms we found that, in addition to the previously described RWD domain in the N-terminal, all these RSUME variants also contain an intermediate domain. Only the longest RSUME isoform presents a C-terminal domain that is absent in the others. Given these differences, we used the shortest and longest RSUME variants for comparative studies. We found that the C-terminal domain is dispensable for the SUMO-conjugation enhancer properties of RSUME. We also demonstrate that these two RSUME variants are equally induced by hypoxia. The NF-κB signaling pathway is inhibited and the HIF-1 pathway is increased more efficiently by the longest RSUME, by means of a greater physical interaction of RSUME267 with the target proteins. In addition, the mRNA and protein levels of these isoforms differ in human glioma samples; while the shortest RSUME isoform is expressed in all the tumors analyzed, the longest variant is expressed in most but not all of them. The results presented here show a degree of redundancy of the RSUME variants on the SUMO pathway. However, the increased inhibition conferred by RSUME267 over the NF-κB signaling pathway, the increased activation over the HIF-1 pathway and the different expression of the RSUME isoforms suggest specific roles for each RSUME isoform which may be relevant in certain types of brain tumors that express RSUME, like human pituitary adenomas and gliomas.
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- 2013
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8. Correction: Corrigendum: Cortical dynamics during cell motility are regulated by CRL3KLHL21 E3 ubiquitin ligase
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Thibault Courtheoux, Radoslav I. Enchev, Fabienne Lampert, Juan Gerez, Jochen Beck, Paola Picotti, Izabela Sumara, and Matthias Peter
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Science - Abstract
Nature Communications 7: Article number: 12810 (2016); Published: 19 September 2016; Updated: 31 October 2016 References cited within the Supplementary Methods section of this Article were not identified in the Supplementary References section. The references are as follows: 44. Enchev, R. I., Schreiber, A.
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- 2016
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9. Detection of cerebral tauopathy in P301L mice using high-resolution large-field multifocal illumination fluorescence microscopy
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Zhenyue Chen, Juan Gerez, Quanyu Zhou, Daniel Razansky, Gloria Shi, Roland Riek, Peter Nilsson, Jan Klohs, Ruiqing Ni, and University of Zurich
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10050 Institute of Pharmacology and Toxicology ,High resolution ,610 Medicine & health ,3107 Atomic and Molecular Physics, and Optics ,01 natural sciences ,Article ,170 Ethics ,010309 optics ,03 medical and health sciences ,0302 clinical medicine ,In vivo ,Atomic and Molecular Physics ,Cortex (anatomy) ,0103 physical sciences ,Microscopy ,Fluorescence microscope ,medicine ,10237 Institute of Biomedical Engineering ,030304 developmental biology ,0303 health sciences ,Chemistry ,medicine.disease ,Fluorescence ,Atomic and Molecular Physics, and Optics ,medicine.anatomical_structure ,1305 Biotechnology ,Biophysics ,Immunohistochemistry ,Tauopathy ,and Optics ,Preclinical imaging ,030217 neurology & neurosurgery ,Intravital microscopy ,Biotechnology - Abstract
Current intravital microscopy techniques visualize tauopathy with high-resolution, but have a small field-of-view and depth-of-focus. Herein, we report a transcranial detection of tauopathy over the entire cortex of P301L tauopathy mice using large-field multifocal illumination (LMI) fluorescence microscopy technique and luminescent conjugated oligothiophenes. In vitro assays revealed that fluorescent ligand h-FTAA is optimal for in vivo tau imaging, which was confirmed by observing elevated probe retention in the cortex of P301L mice compared to non-transgenic littermates. Immunohistochemical staining further verified the specificity of h-FTAA to detect tauopathy in P301L mice. The new imaging platform can be leveraged in pre-clinical mechanistic studies of tau spreading and clearance as well as longitudinal monitoring of tau targeting therapeutics.
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- 2020
10. In-Cell NMR of Intrinsically Disordered Proteins in Mammalian Cells
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Roland Riek, Natalia Cecilia Prymaczok, and Juan Gerez
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0301 basic medicine ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Cell leakage ,Chemistry ,Atomic resolution ,Electroporation ,Biophysics ,α synuclein ,Intrinsically disordered proteins ,Nmr data ,030217 neurology & neurosurgery - Abstract
In-cell NMR enables structural insights at atomic resolution of proteins in their natural environment. To date, very few methods have been developed to study proteins by in-cell NMR in mammalian systems. Here we describe a detailed protocol to conduct in-cell NMR on the intrinsically disordered protein of alpha-Synuclein (αSyn) in mammalian cells. This chapter includes a simplified expression and purification protocol of recombinant αSyn and its delivery into mammalian cells. The chapter also describes how to assess the cell leakage that might occur to the cells, the setup of the instrument, and how to perform basic analyses with the obtained NMR data.
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- 2020
11. Neurodegenerative diseases distinguished through protein-structure analysis
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Roland Riek and Juan Gerez
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0301 basic medicine ,Multidisciplinary ,animal diseases ,Neurodegeneration ,Protein structure analysis ,Disease ,Biology ,medicine.disease ,nervous system diseases ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Atrophy ,nervous system ,mental disorders ,medicine ,Neuroscience ,030217 neurology & neurosurgery - Abstract
Parkinson’s disease and multiple system atrophy involve the protein α-synuclein. Proof that aggregated α-synuclein adopts a different structure in each case suggests that its conformation underlies the distinct disorders. Distinct strains of α-synuclein in two neurodegenerative diseases.
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- 2020
12. A cullin-RING ubiquitin ligase targets exogenous α-synuclein and inhibits Lewy body–like pathology
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Eliezer Masliah, Adriano Aguzzi, Radoslav I. Enchev, Petra Schwarz, Thibault Courtheoux, Matthias Peter, Anthony Adame, Juan Gerez, Paola Picotti, Paul J. Boersema, Uli S. Herrmann, Natalia Cecilia Prymaczok, Roland Riek, Edward Rockenstein, University of Zurich, and Gerez, Juan A
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Pathology ,medicine.medical_specialty ,Proteome ,Ubiquitin-Protein Ligases ,media_common.quotation_subject ,10208 Institute of Neuropathology ,610 Medicine & health ,2700 General Medicine ,Mice ,Neuroblastoma ,03 medical and health sciences ,0302 clinical medicine ,Ubiquitin ,Cell Line, Tumor ,Chlorocebus aethiops ,Skp1 ,medicine ,Animals ,Humans ,Benzothiazoles ,Internalization ,S-Phase Kinase-Associated Proteins ,030304 developmental biology ,media_common ,Neurons ,0303 health sciences ,Lewy body ,biology ,Chemistry ,Parkinson Disease ,General Medicine ,medicine.disease ,Ubiquitin ligase ,COS Cells ,alpha-Synuclein ,biology.protein ,570 Life sciences ,Lewy Bodies ,CUL1 ,030217 neurology & neurosurgery ,Cullin ,Intracellular - Abstract
Parkinson’s disease (PD) is a neurological disorder characterized by the progressive accumulation of neuronal α-synuclein (αSyn) inclusions called Lewy bodies. It is believed that Lewy bodies spread throughout the nervous system due to the cell-to-cell propagation of αSyn via cycles of secretion and uptake. Here, we investigated the internalization and intracellular accumulation of exogenous αSyn, two key steps of Lewy body pathogenesis, amplification and spreading. We found that stable αSyn fibrils substantially accumulate in different cell lines upon internalization, whereas αSyn monomers, oligomers, and dissociable fibrils do not. Our data indicate that the uptake-mediated accumulation of αSyn in a human-derived neuroblastoma cell line triggered an adaptive response that involved proteins linked to ubiquitin ligases of the S-phase kinase-associated protein 1 (SKP1), cullin-1 (Cul1), and F-box domain–containing protein (SCF) family. We found that SKP1, Cul1, and the F-box/LRR repeat protein 5 (FBXL5) colocalized and physically interacted with internalized αSyn in cultured cells. Moreover, the SCF containing the F-box protein FBXL5 (SCF FBXL5 ) catalyzed αSyn ubiquitination in reconstitution experiments in vitro using recombinant proteins and in cultured cells. In the human brain, SKP1 and Cul1 were recruited into Lewy bodies from brainstem and neocortex of patients with PD and related neurological disorders. In both transgenic and nontransgenic mice, intracerebral administration of exogenous αSyn fibrils triggered a Lewy body–like pathology, which was amplified by SKP1 or FBXL5 loss of function. Our data thus indicate that SCF FXBL5 regulates αSyn in vivo and that SCF ligases may constitute targets for the treatment of PD and other α-synucleinopathies.
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- 2019
13. Liquid-liquid phase separation and liquid-to-solid transition mediate α-synuclein amyloid fibril containing hydrogel formation
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Jaladhar Mahato, Rakesh Kumar, Soumik Ray, Sushil Kumar Pandey, Siddhartha Maiti, Ranjith Padinhateeri, Amrendra K. Singh, Samir K. Maji, G. Krishnamoorthy, Juan Gerez, Debalina Datta, Ambuja Navalkar, Laxmikant G. Gadhe, Rajlaxmi Panigrahi, Surabhi Mehra, Ashutosh Kumar, Debdeep Chatterjee, Komal Patel, Nitu Singh, Sandhya Bhatia, Roland Riek, and Arindrajit Chowdhury
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Mutation ,Amyloid ,Chemistry ,animal diseases ,medicine.disease_cause ,In vitro ,nervous system diseases ,Pathogenesis ,Aggresome ,nervous system ,Microtubule ,Biophysics ,medicine ,heterocyclic compounds ,Cellular model ,Oxidative stress - Abstract
SUMMARYα-Synuclein (α-Syn) aggregation and amyloid formation is directly linked with Parkinson’s disease (PD) pathogenesis. However, the early events involved in this process remain unclear. Here, using in vitro reconstitution and cellular model, we show that liquid-liquid phase separation (LLPS) of α-Syn precedes its aggregation. In particular, in vitro generated α-Syn liquid-like droplets eventually undergo a liquid-to-solid transition and form amyloid-hydrogel containing oligomers and fibrillar species. Factors known to aggravate α-Syn aggregation such as low pH, phosphomimic substitution, and familial PD mutation also promote α-Syn LLPS and its subsequent maturation. We further demonstrate α-Syn liquid droplet formation in cells, under oxidative stress. These cellular α-Syn droplets eventually transform into perinuclear aggresomes, the process regulated by microtubules. The present work provides detailed insights into the phase separation behavior of natively unstructured α-Syn and its conversion to a disease-associated aggregated state, which is highly relevant in PD pathogenesis.
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- 2019
14. Cortical dynamics during cell motility are regulated by CRL3KLHL21 E3 ubiquitin ligase
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Jochen Beck, Paola Picotti, Izabela Sumara, Fabienne Lampert, Matthias Peter, Juan Gerez, Radoslav I. Enchev, Thibault Courtheoux, Laboratoire de Biologie Cellulaire et Moléculaire du Contrôle de la Prolifération (LBCMCP), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Centre de Biologie Intégrative (CBI), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institute of Biochemistry (IBC), Eidgenössische Technische Hochschule - Swiss Federal Institute of Technology [Zürich] (ETH Zürich), Institute of Biochemistry, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)
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0301 basic medicine ,Ubiquitin-Protein Ligases ,[SDV]Life Sciences [q-bio] ,Science ,General Physics and Astronomy ,Cell Cycle Proteins ,macromolecular substances ,Biology ,Microtubules ,Article ,General Biochemistry, Genetics and Molecular Biology ,Focal adhesion ,03 medical and health sciences ,Cell Movement ,Cell cortex ,Humans ,Cytoskeleton ,Actin ,ComputingMilieux_MISCELLANEOUS ,Focal Adhesions ,Multidisciplinary ,Ubiquitination ,Cell migration ,General Chemistry ,Corrigenda ,3. Good health ,Cell biology ,Ubiquitin ligase ,Cytoskeletal Proteins ,030104 developmental biology ,biology.protein ,Lamellipodium ,Cortical microtubule ,HeLa Cells - Abstract
Directed cell movement involves spatial and temporal regulation of the cortical microtubule (Mt) and actin networks to allow focal adhesions (FAs) to assemble at the cell front and disassemble at the rear. Mts are known to associate with FAs, but the mechanisms coordinating their dynamic interactions remain unknown. Here we show that the CRL3KLHL21 E3 ubiquitin ligase promotes cell migration by controlling Mt and FA dynamics at the cell cortex. Indeed, KLHL21 localizes to FA structures preferentially at the leading edge, and in complex with Cul3, ubiquitylates EB1 within its microtubule-interacting CH-domain. Cells lacking CRL3KLHL21 activity or expressing a non-ubiquitylatable EB1 mutant protein are unable to migrate and exhibit strong defects in FA dynamics, lamellipodia formation and cortical plasticity. Our study thus reveals an important mechanism to regulate cortical dynamics during cell migration that involves ubiquitylation of EB1 at focal adhesions., Although focal adhesions (FAs) and microtubules (MTs) are known to associate, the underlying regulation of this dynamic interaction is not understood. Here the authors discover that the CRL3KLHL21 E3 ubiquitin ligase localises to FAs and ubiquitinates the MT plus-tip binding protein EB1, thereby promoting MT and FA dynamics and cell migration.
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- 2016
15. Proteomics-Based Monitoring of Pathway Activity Reveals that Blocking Diacylglycerol Biosynthesis Rescues from Alpha-Synuclein Toxicity
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Paul J. Boersema, Martin Soste, Matthias Peter, Stefano Vanni, Juan Gerez, Konstantina Charmpi, Roland Riek, Andreas Beyer, Paola Picotti, Marc van Oostrum, Liliana Malinovska, Fabienne Lampert, and Natalia Cecilia Prymaczok
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Proteomics ,Histology ,Saccharomyces cerevisiae Proteins ,Phosphoproteomics ,animal diseases ,Phosphatidate Phosphatase ,Apoptosis ,Saccharomyces cerevisiae ,Protein aggregation ,Article ,Pathology and Forensic Medicine ,Alpha-synuclein ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Pah1 ,Lipid droplet ,Gene Expression Regulation, Fungal ,Humans ,Molecular Targeted Therapy ,030304 developmental biology ,Diacylglycerol kinase ,Neurons ,0303 health sciences ,Lipin ,Chemistry ,Effector ,Endoplasmic reticulum ,Galactolipids ,Lipid metabolism ,Parkinson Disease ,Cell Biology ,Phosphatidic acid ,Lipid Droplets ,Lipid Metabolism ,3. Good health ,Cell biology ,nervous system diseases ,Sentinel proteins ,nervous system ,Toxicity ,Parkinson’s disease ,Diacylglycerol ,030217 neurology & neurosurgery ,Signal Transduction - Abstract
Summary Proteinaceous inclusions containing alpha-synuclein (α-Syn) have been implicated in neuronal toxicity in Parkinson’s disease, but the pathways that modulate toxicity remain enigmatic. Here, we used a targeted proteomic assay to simultaneously measure 269 pathway activation markers and proteins deregulated by α-Syn expression across a panel of 33 Saccharomyces cerevisiae strains that genetically modulate α-Syn toxicity. Applying multidimensional linear regression analysis to these data predicted Pah1, a phosphatase that catalyzes conversion of phosphatidic acid to diacylglycerol at the endoplasmic reticulum membrane, as an effector of rescue. Follow-up studies demonstrated that inhibition of Pah1 activity ameliorates the toxic effects of α-Syn, indicate that the diacylglycerol branch of lipid metabolism could enhance α-Syn neuronal cytotoxicity, and suggest a link between α-Syn toxicity and the biology of lipid droplets., Graphical Abstract, Highlights • Proteomics of pathway sentinels across 33 genetic modulators of α-Synuclein toxicity • Multidimensional linear regression predicts lipid metabolism to regulate rescue • Genetic and chemical perturbation of Pah1 alters α-Synuclein inclusions and toxicity • Diacylglycerol and lipid droplets may play a role in cytotoxicity induced by α-Synuclein, α-Synuclein is genetically and neuropathologically linked to Parkinson’s disease. However, the mechanisms of known genetic toxicity modulators in a yeast model system remain largely unknown. In order to identify cellular rescue pathways at high-throughput, we have paired mass spectrometry-based monitoring of pathway activity and growth profiles through regression analysis. The results predicted a critical role for the protein Pah1 in lipid metabolism. Indeed, specific perturbation of Pah1 activity determines inclusion formation and toxicity thereby suggesting a potential target for combating pathologies associated with α-Synuclein accumulation.
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- 2018
16. RSUME inhibits VHL and regulates its tumor suppressor function
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Marcelo Paez-Pereda, Mariana Fuertes, G. K. Stalla, M Barontini, Lucas Tedesco, Florian Holsboer, Ulrich Renner, Eduardo Arzt, Yonghe Wu, Juan Gerez, Susana Silberstein, and Juan José Bonfiglio
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Male ,Cancer Research ,medicine.medical_specialty ,CIENCIAS MÉDICAS Y DE LA SALUD ,Adrenal Gland Neoplasms ,Down-Regulation ,Mice, Nude ,Pheochromocytoma ,Pvhl ,Biology ,law.invention ,Mice ,law ,Internal medicine ,Chlorocebus aethiops ,Tumor Cells, Cultured ,Genetics ,medicine ,Hif ,Animals ,Humans ,Genes, Tumor Suppressor ,Von Hippel–Lindau disease ,Hypoxia ,Cerebellar Neoplasms ,Molecular Biology ,Ubiquitination ,Bioquímica y Biología Molecular ,Rwdd3 ,medicine.disease ,Hemangioblastoma ,Gene Expression Regulation, Neoplastic ,Medicina Básica ,Endocrinology ,Von Hippel-Lindau Tumor Suppressor Protein ,COS Cells ,Disease Progression ,Cancer research ,Von Hippel-Lindau Disease ,Suppressor ,Function (biology) ,Transcription Factors - Abstract
Somatic mutations or loss of von Hippel-Lindau (pVHL) happen in the majority of VHL disease tumors, which present a constitutively active Hypoxia Inducible Factor (HIF), essential for tumor growth. Recently described mechanisms for pVHL modulation shed light on the open question of the HIF/pVHL pathway regulation. The aim of the present study was to determine the molecular mechanism by which RSUME stabilizes HIFs, by studying RSUME effect on pVHL function and to determine the role of RSUME on pVHL-related tumor progression. We determined that RSUME sumoylates and physically interacts with pVHL and negatively regulates the assembly of the complex between pVHL, Elongins and Cullins (ECV), inhibiting HIF-1 and 2alpha ubiquitination and degradation. We found that RSUME is expressed in human VHL tumors (renal clear-cell carcinoma (RCC), pheochromocytoma and hemangioblastoma) and by overexpressing or silencing RSUME in a pVHL-HIF-oxygen-dependent degradation stability reporter assay, we determined that RSUME is necessary for the loss of function of type 2 pVHL mutants. The functional RSUME/pVHL interaction in VHL-related tumor progression was further confirmed using a xenograft assay in nude mice. RCC clones, in which RSUME was knocked down and express either pVHL wt or type 2 mutation, have an impaired tumor growth, as well as HIF-2alpha, vascular endothelial growth factor A and tumor vascularization diminution. This work shows a novel mechanism for VHL tumor progression and presents a new mechanism and factor for targeting tumor-related pathologies with pVHL/HIF altered function. Fil: Gerez, Juan Atilio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina Fil: Tedesco, Lucas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina Fil: Bonfiglio, Juan José. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina Fil: Fuertes, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina Fil: Barontini, Marta Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Centro de Investigaciones Endocrinológicas "Dr. César Bergada". Gobierno de la Ciudad de Buenos Aires. Centro de Investigaciones Endocrinológicas "Dr. César Bergada". Fundación de Endocrinología Infantil. Centro de Investigaciones Endocrinológicas "Dr. César Bergada"; Argentina Fil: Silberstein Cuña, Susana Iris. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina Fil: Wu, Y.. Max Planck Institute of Psychiatry; Alemania Fil: Renner, U.. Max Planck Institute of Psychiatry; Alemania Fil: Paez Pereda, M.. Max Planck Institute of Psychiatry; Alemania Fil: Holsboer, F.. Max Planck Institute of Psychiatry; Alemania Fil: Stalla, G. K.. Max Planck Institute of Psychiatry; Alemania Fil: Arzt, Eduardo Simon. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina
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- 2014
17. Regulation of α-synuclein by chaperones in mammalian cells
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Thomas Bock, Darius Šulskis, Roland Riek, Silvia Campioni, Dhiman Ghosh, Juan Gerez, Adam Mazur, Björn M. Burmann, Irena Matečko-Burmann, Stefan G.D. Rüdiger, Magdalena Wawrzyniuk, Pratibha Kumari, Sebastian Hiller, Alexander Schmidt, Emelie E. Aspholm, Immunologie, Dep Scheikunde, and Sub Cellular Protein Chemistry
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0301 basic medicine ,Magnetic Resonance Spectroscopy ,Cell Survival ,Mitochondrion ,C-abl ,Article ,03 medical and health sciences ,0302 clinical medicine ,Complex ,Taverne ,medicine ,Humans ,Forms ,Conformation ,Multidisciplinary ,biology ,Chemistry ,Kinase ,Reveals ,HEK 293 cells ,Neurodegeneration ,Binding ,Skp ,medicine.disease ,Hsp90 ,Cell biology ,nervous system diseases ,030104 developmental biology ,HEK293 Cells ,Chaperone (protein) ,biology.protein ,Specificity ,alpha-Synuclein ,Phosphorylation ,Protein Processing, Post-Translational ,030217 neurology & neurosurgery ,Intracellular ,Molecular Chaperones - Abstract
Neurodegeneration in patients with Parkinson's disease is correlated with the occurrence of Lewy bodies-intracellular inclusions that contain aggregates of the intrinsically disordered protein α-synuclein; 1; . The aggregation propensity of α-synuclein in cells is modulated by specific factors that include post-translational modifications; 2,3; , Abelson-kinase-mediated phosphorylation; 4,5; and interactions with intracellular machineries such as molecular chaperones, although the underlying mechanisms are unclear; 6-8; . Here we systematically characterize the interaction of molecular chaperones with α-synuclein in vitro as well as in cells at the atomic level. We find that six highly divergent molecular chaperones commonly recognize a canonical motif in α-synuclein, consisting of the N terminus and a segment around Tyr39, and hinder the aggregation of α-synuclein. NMR experiments; 9; in cells show that the same transient interaction pattern is preserved inside living mammalian cells. Specific inhibition of the interactions between α-synuclein and the chaperone HSC70 and members of the HSP90 family, including HSP90β, results in transient membrane binding and triggers a remarkable re-localization of α-synuclein to the mitochondria and concomitant formation of aggregates. Phosphorylation of α-synuclein at Tyr39 directly impairs the interaction of α-synuclein with chaperones, thus providing a functional explanation for the role of Abelson kinase in Parkinson's disease. Our results establish a master regulatory mechanism of α-synuclein function and aggregation in mammalian cells, extending the functional repertoire of molecular chaperones and highlighting new perspectives for therapeutic interventions for Parkinson's disease.
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- 2017
18. Accumulation of oligomer-prone alpha-synuclein exacerbates synaptic and neuronal degeneration in vivo
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Eliezer Masliah, Paola Picotti, Christina Patrick, Fred H. Gage, Margarita Trejo-Morales, Jürgen Winkler, Michael Mante, Edward Rockenstein, Anthony Adame, Kiren Ubhi, Cassia R. Overk, Poul Henning Jensen, Silke Nuber, Silvia Campioni, Beate Winner, Roland Riek, and Juan Gerez
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animal diseases ,Hippocampus ,Synapse ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,Medizinische Fakultät ,Antigens, Thy-1 ,Neurons ,0303 health sciences ,Neurodegeneration ,Brain ,3. Good health ,Cell biology ,alpha-Synuclein ,Genetically modified mouse ,Lewy Body Disease ,Synapsin I ,Glutamic Acid ,Mice, Transgenic ,Nerve Tissue Proteins ,Biology ,Synaptic vesicle ,03 medical and health sciences ,Alzheimer Disease ,mental disorders ,medicine ,Animals ,Humans ,ddc:610 ,030304 developmental biology ,Alpha-synuclein ,Memory Disorders ,Dementia with Lewy bodies ,Lysine ,Original Articles ,Transgenic ,Oligomer ,Parkinson's disease ,Synaptic vesicles ,medicine.disease ,nervous system diseases ,Mice, Inbred C57BL ,Disease Models, Animal ,chemistry ,nervous system ,Gene Expression Regulation ,Mutation ,Nerve Degeneration ,Synapses ,Thy-1 Antigens ,Neurology (clinical) ,Neuroscience ,030217 neurology & neurosurgery - Abstract
In Parkinson's disease and dementia with Lewy bodies, α-synuclein aggregates to form oligomers and fibrils; however, the precise nature of the toxic α-synuclein species remains unclear. A number of synthetic α-synuclein mutations were recently created (E57K and E35K) that produce species of α-synuclein that preferentially form oligomers and increase α-synuclein-mediated toxicity. We have shown that acute lentiviral expression of α-synuclein E57K leads to the degeneration of dopaminergic neurons; however, the effects of chronic expression of oligomer-prone α-synuclein in synapses throughout the brain have not been investigated. Such a study could provide insight into the possible mechanism(s) through which accumulation of α-synuclein oligomers in the synapse leads to neurodegeneration. For this purpose, we compared the patterns of neurodegeneration and synaptic damage between a newly generated mThy-1 α-synuclein E57K transgenic mouse model that is prone to forming oligomers and the mThy-1 α-synuclein wild-type mouse model (Line 61), which accumulates various forms of α-synuclein. Three lines of α-synuclein E57K (Lines 9, 16 and 54) were generated and compared with the wild-type. The α-synuclein E57K Lines 9 and 16 were higher expressings of α-synuclein, similar to α-synuclein wild-type Line 61, and Line 54 was a low expressing of α-synuclein compared to Line 61. By immunoblot analysis, the higher-expressing α-synuclein E57K transgenic mice showed abundant oligomeric, but not fibrillar, α-synuclein whereas lower-expressing mice accumulated monomeric α-synuclein. Monomers, oligomers, and fibrils were present in α-synuclein wild-type Line 61. Immunohistochemical and ultrastructural analyses demonstrated that α-synuclein accumulated in the synapses but not in the neuronal cells bodies, which was different from the α-synuclein wild-type Line 61, which accumulates α-synuclein in the soma. Compared to non-transgenic and lower-expressing mice, the higher-expressing α-synuclein E57K mice displayed synaptic and dendritic loss, reduced levels of synapsin 1 and synaptic vesicles, and behavioural deficits. Similar alterations, but to a lesser extent, were seen in the α-synuclein wild-type mice. Moreover, although the oligomer-prone α-synuclein mice displayed neurodegeneration in the frontal cortex and hippocampus, the α-synuclein wild-type only displayed neuronal loss in the hippocampus. These results support the hypothesis that accumulating oligomeric α-synuclein may mediate early synaptic pathology in Parkinson's disease and dementia with Lewy bodies by disrupting synaptic vesicles. This oligomer-prone model might be useful for evaluating therapies directed at oligomer reduction.
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- 2014
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19. RSUME Enhances Glucocorticoid Receptor SUMOylation and Transcriptional Activity
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Eduardo Arzt, Jimena Druker, Jorge A. Iñiguez-Lluhí, Florian Holsboer, Ana C. Liberman, María Antunica-Noguerol, Juan Gerez, Marcelo Paez-Pereda, and Theo Rein
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Transcriptional Activation ,Protein sumoylation ,Lysine ,SUMO protein ,Nerve Tissue Proteins ,Arginine ,Cell Line ,Receptors, Glucocorticoid ,Glucocorticoid receptor ,Ubiquitin ,Chlorocebus aethiops ,Coactivator ,Animals ,Enhancer ,Molecular Biology ,Transcription factor ,biology ,Sumoylation ,Articles ,Cell Biology ,Molecular biology ,Protein Structure, Tertiary ,Rats ,COS Cells ,Mutation ,biology.protein ,Carrier Proteins ,Heat-Shock Response ,Transcription Factors - Abstract
Glucocorticoid receptor (GR) activity is modulated by posttranslational modifications, including phosphorylation, ubiquitination, and SUMOylation. The GR has three SUMOylation sites: lysine 297 (K297) and K313 in the N-terminal domain (NTD) and K721 within the ligand-binding domain. SUMOylation of the NTD sites mediates the negative effect of the synergy control motifs of GR on promoters with closely spaced GR binding sites. There is scarce evidence on the role of SUMO conjugation to K721 and its impact on GR transcriptional activity. We have previously shown that RSUME (RWD-containing SUMOylation enhancer) increases protein SUMOylation. We now demonstrate that RSUME interacts with the GR and increases its SUMOylation. RSUME regulates GR transcriptional activity and the expression of its endogenous target genes, FKBP51 and S100P. RSUME uncovers a positive role for the third SUMOylation site, K721, on GR-mediated transcription, demonstrating that GR SUMOylation acts positively in the presence of a SUMOylation enhancer. Both mutation of K721 and small interfering RNA-mediated RSUME knockdown diminish GRIP1 coactivator activity. RSUME, whose expression is induced under stress conditions, is a key factor in heat shock-induced GR SUMOylation. These results show that inhibitory and stimulatory SUMO sites are present in the GR and at higher SUMOylation levels the stimulatory one becomes dominant.
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- 2013
20. Co-Expression of Wild-Type P2X7R with Gln460Arg Variant Alters Receptor Function
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Matías Acuña, Florian Holsboer, Sergio A. Senin, Miso Mitkovski, Herbert Stadler, Markus Panhuysen, Marcelo Paez-Pereda, Jan M. Deussing, Esteban Hoijman, Ana C. Liberman, Eduardo Arzt, Juan Gerez, Damian Refojo, Walter Stühmer, Michael W. Metzger, and Fernando Aprile-Garcia
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0301 basic medicine ,Patch-Clamp Techniques ,Physiology ,Cell Membranes ,lcsh:Medicine ,Artificial Gene Amplification and Extension ,Biochemistry ,Polymerase Chain Reaction ,Fluorophotometry ,Intracellular Receptors ,Ion Channels ,0302 clinical medicine ,Spectrum Analysis Techniques ,Fluorescence Resonance Energy Transfer ,Medicine and Health Sciences ,Small interfering RNAs ,RNA, Small Interfering ,lcsh:Science ,Receptor ,Multidisciplinary ,Voltage-dependent calcium channel ,Physics ,Purinergic receptor ,purl.org/becyt/ford/3.1 [https] ,Bioquímica y Biología Molecular ,Gln460arg ,Px27r ,Nucleic acids ,Electrophysiology ,Medicina Básica ,Spectrophotometry ,Physical Sciences ,Ligand-gated ion channel ,purl.org/becyt/ford/3 [https] ,medicine.symptom ,Cellular Structures and Organelles ,Signal Transduction ,Research Article ,CIENCIAS MÉDICAS Y DE LA SALUD ,Blotting, Western ,Biophysics ,Heterodimerization ,Neurophysiology ,Biology ,Real-Time Polymerase Chain Reaction ,Research and Analysis Methods ,Polymorphism, Single Nucleotide ,03 medical and health sciences ,medicine ,Genetics ,Humans ,Immunoprecipitation ,Non-coding RNA ,Molecular Biology Techniques ,Molecular Biology ,Loss function ,Biology and life sciences ,lcsh:R ,Wild type ,Proteins ,P2RX7 ,Cell Biology ,Ligand-Gated Ion Channels ,Molecular biology ,Gene regulation ,030104 developmental biology ,HEK293 Cells ,Mechanism of action ,RNA ,lcsh:Q ,Calcium ,Receptors, Purinergic P2X7 ,Gene expression ,Calcium Channels ,030217 neurology & neurosurgery ,Cloning ,Neuroscience - Abstract
The P2X7 receptor is a member of the P2X family of ligand-gated ion channels. A single-nucleotide polymorphism leading to a glutamine (Gln) by arginine (Arg) substitution at codon 460 of the purinergic P2X7 receptor (P2X7R) has been associated with mood disorders. No change in function (loss or gain) has been described for this SNP so far. Here we show that although the P2X7R-Gln460Arg variant per se is not compromised in its function, co-expression of wild-type P2X7R with P2X7R-Gln460Arg impairs receptor function with respect to calcium influx, channel currents and intracellular signaling in vitro. Moreover, co-immunoprecipitation and FRET studies show that the P2X7R-Gln460Arg variant physically interacts with P2X7R-WT. Specific silencing of either the normal or polymorphic variant rescues the heterozygous loss of function phenotype and restores normal function. The described loss of function due to co-expression, unique for mutations in the P2RX7 gene so far, explains the mechanism by which the P2X7R-Gln460Arg variant affects the normal function of the channel and may represent a mechanism of action for other mutations., PLoS ONE, 11 (3), ISSN:1932-6203
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- 2015
21. Intrapituitary Expression and Regulation of the gp130 Cytokine Interleukin-6 and Its Implication in Pituitary Physiology and Pathophysiology
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Marcelo Paez Pereda, Eliane Correa De Santana, Günter K. Stalla, Mariana Haedo, Bianca Fröhlich, Juan Gerez, Eduardo Arzt, C. Onofri, and Ulrich Renner
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medicine.medical_specialty ,medicine.medical_treatment ,Biology ,General Biochemistry, Genetics and Molecular Biology ,History and Philosophy of Science ,Anterior pituitary ,Internal medicine ,Cytokine Receptor gp130 ,medicine ,Animals ,Humans ,Pituitary Neoplasms ,Interleukin 6 ,Innate immune system ,Interleukin-6 ,General Neuroscience ,Pituitary tumors ,Interleukin ,medicine.disease ,Glycoprotein 130 ,Cytokine ,Endocrinology ,medicine.anatomical_structure ,Pituitary Gland ,biology.protein ,Corticotropic cell - Abstract
Interleukin (IL)-6, a member of the gp130 cytokine family, is sometimes designated as an "endocrine" cytokine because of its strong regulatory influence on hormone production. Systemically acting IL-6 derived from immune cells is a potent stimulator of the hypothalamus-pituitary-adrenal axis and therefore plays an important role in modulating immune-neuroendocrine interactions during inflammatory or infectious processes. However, IL-6 is also produced within the anterior pituitary by so-called folliculostellate (FS) cells and is also synthesized in and released by tumor cells in pituitary adenomas. Growth factors (e.g., transforming growth factor-beta), neuropeptides (e.g., pituitary adenylate cyclase-activating polypeptide), or hormones (e.g., glucocorticoids) regulate IL-6 production both in FS and pituitary tumor cells. Interestingly, components of the innate immune system, such as toll-like receptor 4 and nucleotide-binding oligomerization domains (NODs), are expressed in FS and pituitary tumor cells. Therefore, cell-wall components of bacteria (lipopolysaccharide, muramyl dipeptide, diamino pimelic acid) stimulate IL-6 production in normal and tumoral pituitary. The intrinsic IL-6 production by FS cells in normal anterior pituitary may participate in immune-neuroendocrine interactions during inflammatory processes. In pituitary adenomas, IL-6 stimulates hormone secretion, tumor cell proliferation, and the production of angiogenic factors, such as vascular endothelial growth factor-A, suggesting an important role of IL-6 in the pathophysiology and progression of pituitary adenomas.
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- 2009
22. Differential Gene Expression in Models of Pituitary Prolactin-Producing Tumoral Cells
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Damiana Giacomini, Günter K. Stalla, Eduardo Arzt, Juan Gerez, Marcelo Paez-Pereda, Marta Labeur, Jimena Druker, and Mariana Haedo
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medicine.medical_specialty ,Endocrinology, Diabetes and Metabolism ,Bone Morphogenetic Protein 4 ,macromolecular substances ,Biology ,Models, Biological ,Mice ,Endocrinology ,Cell Line, Tumor ,Internal medicine ,TGF beta signaling pathway ,Gene expression ,medicine ,Animals ,Humans ,Prolactinoma ,Gene ,Pituitary tumours ,Mice, Knockout ,Receptors, Dopamine D2 ,Effector ,Gene Expression Profiling ,Cushing's disease ,medicine.disease ,Cell Hypoxia ,Prolactin ,Neoplasm Proteins ,Gene Expression Regulation, Neoplastic ,Cell Transformation, Neoplastic ,Pediatrics, Perinatology and Child Health ,Cancer research ,Signalling pathways ,Signal Transduction ,Transcription Factors - Abstract
Although several genes and signalling pathways have been identified as important effectors in the development of pituitary tumours, our understanding of pituitary tumorigenesis remains incomplete and is the focus of much current research. Use of the mRNA differential display technique in prolactinomas from D2-receptor knockout mice and in stable GH3 cell line clones with enhanced tumorigenicity in vivo has led to the identification of two genes that are involved in the pathogenic process – BMP-4 and RSUME. Bone morphogenetic protein-4 (BMP-4) has been found to have a crucial role in prolactinoma development and also in signalling crosstalk with oestrogens. In contrast, BMP-4 has an inhibitory role in corticotrophinomas. RSUME (RWD-containing sumoylation enhancer) was identified from a transformed lactosomatotrophic cell line that had increased tumorigenic and angiogenic potential. Expression of RSUME was induced under hypoxic conditions and it has a potential role during vascularization. The differential expression and action of BMP-4 in prolactinomas and corticotrophinomas highlights the importance of studying a gene with contrasting actions in two cell lineages of the same organ in order to understand the pituitary transformation process. Both BMP-4 and RSUME may be interesting targets for inhibiting steps involved in pituitary tumorigenesis.
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- 2009
23. Regulation of Pituitary Function by Cytokines
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Damiana Giacomini, Eduardo Arzt, Ulrich Renner, Mariana Fuertes, Mariana Haedo, Marta Labeur, Juan Gerez, Marcelo Paez-Pereda, and Günter K. Stalla
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Adenoma ,medicine.medical_specialty ,Pituitary gland ,Somatotropic cell ,Endocrinology, Diabetes and Metabolism ,medicine.medical_treatment ,Pituitary neoplasm ,Biology ,Models, Biological ,Prolactin cell ,Endocrinology ,Internal medicine ,medicine ,Animals ,Humans ,Pituitary Neoplasms ,Cellular Senescence ,Interleukin-6 ,Pituitary tumors ,medicine.disease ,Cytokine ,medicine.anatomical_structure ,Pituitary Gland ,Pediatrics, Perinatology and Child Health ,Cytokines ,Cell aging - Abstract
Research performed on the pituitary has proven that cytokines play an important role in maintaining pituitary physiology, affecting not only cell proliferation but also hormone secretion. The effects of cytokines can be autocrine or paracrine. This review gives an overview on the effects of the most studied cytokines in the pituitary. Special interest is focused on interleukin-6 (IL-6) because it has the distinctive characteristic of stimulating pituitary tumor cell growth, but has the opposite effect on normal pituitary cells. On the other hand, IL-6 is a cytokine of interest in the pituitary because recent work has shown that it promotes and maintains senescence in certain types of tumors. Given that the majority of pituitary adenomas are microadenomas and the fact that clinically inapparent pituitary tumors are quite common, senescence, perhaps mediated by IL-6, is an attractive mechanism for explaining the benign nature of pituitary tumors.
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- 2009
24. RSUME, a Small RWD-Containing Protein, Enhances SUMO Conjugation and Stabilizes HIF-1α during Hypoxia
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Susana Silberstein, Eduardo Arzt, Florian Holsboer, Günter K. Stalla, Alberto Carbia-Nagashima, Juan Gerez, Carolina Perez-Castro, and Marcelo Paez-Pereda
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SUMO-1 Protein ,SUMO protein ,animal cell ,protein binding ,Plasma protein binding ,Ubiquitin-conjugating enzyme ,I kappa B ,Chlorocebus aethiops ,rat ,sumoylation ,article ,NF-kappa B ,protein domain ,sumo 2 protein ,Cell Hypoxia ,unclassified drug ,I-kappa B Kinase ,Cell biology ,immunoglobulin enhancer binding protein ,priority journal ,protein stability ,Biochemistry ,Organ Specificity ,polymerization ,protein protein interaction ,Mammalia ,Small Ubiquitin-Related Modifier Proteins ,conjugation ,Protein Binding ,regulatory mechanism ,PROTEINS ,Molecular Sequence Data ,Protein domain ,protein Ubc9 ,macromolecular substances ,Biology ,General Biochemistry, Genetics and Molecular Biology ,Cercopithecus aethiops ,Protein–protein interaction ,Cell Line, Tumor ,Animals ,Humans ,controlled study ,hypoxia inducible factor 1alpha ,human ,Amino Acid Sequence ,cell protein ,protein structure ,protein expression ,Ubiquitins ,Transcription factor ,mouse ,nonhuman ,Base Sequence ,hypoxia ,Biochemistry, Genetics and Molecular Biology(all) ,human cell ,genetic transcription ,nucleotide sequence ,protein RSUME ,Hypoxia-Inducible Factor 1, alpha Subunit ,sumo 3 protein ,Ubiquitin-Conjugating Enzymes ,cell function ,CELLBIO ,mammal cell ,SUMO 1 protein ,Transcription Factors - Abstract
SUMO conjugation to proteins is involved in the regulation of diverse cellular functions. We have identified a protein, RWD-containing sumoylation enhancer (RSUME), that enhances overall SUMO-1, -2, and -3 conjugation by interacting with the SUMO conjugase Ubc9. RSUME increases noncovalent binding of SUMO-1 to Ubc9 and enhances Ubc9 thioester formation and SUMO polymerization. RSUME enhances the sumoylation of IkB in vitro and in cultured cells, leading to an inhibition of NF-kB transcriptional activity. RSUME is induced by hypoxia and enhances the sumoylation of HIF-1α, promoting its stabilization and transcriptional activity during hypoxia. Disruption of the RWD domain structure of RSUME demonstrates that this domain is critical for RSUME action. Together, these findings point to a central role of RSUME in the regulation of sumoylation and, hence, several critical regulatory pathways in mammalian cells. © 2007 Elsevier Inc. All rights reserved. Fil:Carbia-Nagashima, A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Gerez, J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Perez-Castro, C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Paez-Pereda, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Silberstein, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
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- 2007
25. Molecular transduction mechanisms of cytokine-hormone interactions: role of gp130 cytokines
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Juan Gerez, José Bonfiglio, Damiana Giacomini, Marcelo J. Perone, Ulrich Renner, Sebastian Sosa, Alberto Carbia Nagashima, Günter K. Stalla, Eduardo Arzt, Matias Acuña, and Susana Silberstein
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Transduction (genetics) ,Immune system ,Cytokine ,Chemistry ,medicine.medical_treatment ,medicine ,General Medicine ,Glycoprotein 130 ,Hormone ,Cell biology - Published
- 2007
26. Molecular transduction mechanisms of cytokine hormone interactions: role of gp130 cytokines
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Günter K. Stalla, Susana Silberstein, Damiana Giacomini, José Bonfiglio, Eduardo Arzt, Ulrich Renner, Sebastian Sosa, Marcelo J. Perone, Matias Acuña, Alberto Carbia Nagashima, and Juan Gerez
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medicine.medical_treatment ,General Medicine ,Biology ,Glycoprotein 130 ,Therapeutic approach ,Transduction (genetics) ,Cytokine ,medicine.anatomical_structure ,Immune system ,Anterior pituitary ,Immunology ,medicine ,Receptor ,Hormone - Abstract
Highly sophisticated mechanisms confer on the immune system the capacity to respond with a certain degree of autonomy. However, the final outcome of an immune response depends on the interaction of the immune system with other systems. The immune and neuroendocrine systems have an intimate cross-communication that makes possible a satisfactory response to environmental changes. Part of this interaction occurs through cytokines and steroid hormones. The last step of this cross-talk is the molecular level. As a model of interaction, this review focuses on the gp130 cytokine family. These cytokines, as well as their receptors, are expressed in pituitary cells. They regulate hormone production as well as growth of pituitary cells. During acute or chronic inflammation or infection, systemic, hypothalamic and hypophyseal gp130 cytokines act on anterior pituitary cells, integrating the neuroendocrine–immune response. Disruptions of these pathways may lead not only to abnormal growth of pituitary cells but also to immune disorders, for which, based on recent findings, targeting these cytokines might be a novel therapeutic approach.
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- 2007
27. Correction: Corrigendum: Cortical dynamics during cell motility are regulated by CRL3KLHL21 E3 ubiquitin ligase
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Jochen Beck, Matthias Peter, Thibault Courtheoux, Fabienne Lampert, Paola Picotti, Juan Gerez, Radoslav I. Enchev, and Izabela Sumara
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0301 basic medicine ,Multidisciplinary ,biology ,Published Erratum ,Science ,Section (typography) ,General Physics and Astronomy ,Motility ,General Chemistry ,General Biochemistry, Genetics and Molecular Biology ,Cell biology ,Ubiquitin ligase ,03 medical and health sciences ,030104 developmental biology ,biology.protein - Abstract
Nature Communications 7: Article number: 12810 (2016); Published: 19 September 2016; Updated: 31 October 2016 References cited within the Supplementary Methods section of this Article were not identified in the Supplementary References section. The references are as follows: 44. Enchev, R. I., Schreiber, A.
- Published
- 2016
28. RSUME is implicated in HIF-1-induced VEGF-A production in pituitary tumour cells
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Juan Gerez, Mariana Fuertes, Günter K. Stalla, Mariana Haedo, Marco Losa, Eduardo Arzt, Bing Shan, Ulrich Renner, Michael Buchfelder, Marily Theodoropoulou, Shan, B., Gerez, J., Haedo, M., Fuertes, M., Theodoropoulou, M., Buchfelder, M., Losa, M., Stalla, G. K., Arzt, E., and Renner, U.
- Subjects
Adenoma ,Vascular Endothelial Growth Factor A ,Cancer Research ,medicine.medical_specialty ,CIENCIAS MÉDICAS Y DE LA SALUD ,Transcription Factor ,Endocrinology, Diabetes and Metabolism ,Biology ,Pituitary neoplasm ,Mice ,Endocrinology ,Pituitary adenoma ,Cell Line, Tumor ,Internal medicine ,Tumor Cells, Cultured ,medicine ,Animals ,Humans ,Pituitary Neoplasms ,Secretion ,Pituitary Neoplasm ,RNA, Messenger ,Hypoxia ,Transcription factor ,Messenger RNA ,Neovascularization, Pathologic ,Animal ,Pituitary tumour ,Cobalt ,Hypoxia-Inducible Factor 1, alpha Subunit ,medicine.disease ,Otras Ciencias Médicas ,Vascular endothelial growth factor A ,Oncology ,Cell culture ,Cancer research ,RSUME ,Transcription Factors ,Human - Abstract
The recently cloned small RWD-domain containing protein RSUME was shown to increase protein levels of hypoxia-inducible factor-1a (HIF-1a). The latter is the oxygen-regulated subunit of HIF-1, the most important transcription factor of the cellular adaptive processes to hypoxic conditions. It is also a major regulator of vascular endothelial growth factor-A (VEGF-A), which is critically involved in the complex process of tumour neovascularisation. In this study, the expression and role of RSUME in pituitary tumours was studied. We found that RSUME mRNA was up-regulated in pituitary adenomas and significantly correlated with HIF-1a mRNA levels. Hypoxia (1% O2) or treatment with hypoxia-mimicking CoCl2 enhanced RSUME and HIF-1a expression, induced translocation of HIF-1a to the nuclei and stimulated VEGF-A production both in pituitary tumour cell lines and primary human pituitary adenoma cell cultures. When RSUME expression was specifically down-regulated by siRNA, the CoCl2-induced increase VEGF-A secretion was strongly reduced which was shown to be a consequence of the RSUME knockdown-associated reduction of HIF-1a synthesis. Thus, RSUME plays an important role in initiating pituitary tumour neovascularisation through regulating HIF-1a levels and subsequent VEGF-A production and may therefore be critically involved in pituitary adenoma progression. Fil: Shan, B.. Max-Planck-Institut für Psychiatrie; Alemania Fil: Gerez, Juan Atilio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Pque. Centenario. Instituto de Investigación En Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina Fil: Haedo, Mariana Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Pque. Centenario. Instituto de Investigación En Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina Fil: Fuertes, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Pque. Centenario. Instituto de Investigación En Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina Fil: Theodoropoulou, M.. Max-Planck-Institut für Psychiatrie; Alemania Fil: Buchfelder, M.. Universitat Erlangen-Nuremberg; Alemania Fil: Losa, M.. Istituto San Raffaele; Italia Fil: Stalla, G.. Max-Planck-Institut für Psychiatrie; Alemania Fil: Arzt, Eduardo Simon. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Pque. Centenario. Instituto de Investigación En Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina Fil: Renner, U.. Max-Planck-Institut für Psychiatrie; Alemania
- Published
- 2012
29. The Effects of Hypoxia-Inducible Factor on Growth Hormone in GH3 Cells
- Author
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Mariana Raquel Haedo, Mariana Fuertes, Juan Gerez, Ulrich Renner, Johanna Stalla, Gunter K Stalla, and Eduardo Arzt
- Published
- 2011
30. The serine/arginine-rich protein SF2/ASF regulates protein sumoylation
- Author
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Federico Pelisch, Eduardo Arzt, Belinda J. Westman, Guillermo Risso, Jimena Druker, Anabella Srebrow, Ezequiel Petrillo, Juan Gerez, Angus I. Lamond, Ignacio E. Schor, and Manuel Javier Muñoz
- Subjects
RNA splicing ,protein p53 ,SUMO protein ,protein binding ,Heterogeneous ribonucleoprotein particle ,environment and public health ,Substrate Specificity ,nuclear protein ,Transcriptional regulation ,genetics ,RNA, Small Interfering ,E3 ligase ,serine-arginine-rich splicing proteins ,Multidisciplinary ,protein asf ,biology ,Serine-Arginine Splicing Factors ,sumoylation ,article ,protein processing ,Nuclear Proteins ,RNA-Binding Proteins ,ubiquitin-conjugating enzyme UBC9 ,cell line ,protein function ,Biological Sciences ,heat shock ,unclassified drug ,enzyme activity ,Ubiquitin ligase ,ubiquitin conjugating enzyme ,regulator protein ,priority journal ,Biochemistry ,protein protein interaction ,ubiquitin protein ligase E3 ,signal transduction ,conjugation ,Protein Binding ,Protein sumoylation ,regulatory mechanism ,SUMO-1 Protein ,protein Ubc9 ,SUMO enzymes ,Cell Line ,heat shock response ,SR protein ,Humans ,controlled study ,human ,enzyme specificity ,protein expression ,protein inhibitor of activated STAT1 ,RNA metabolism ,Splicing factor ,protein inhibitor of activated STAT ,human cell ,SUMO E3 ligase ,RNA binding protein ,small interfering RNA ,Sumoylation Pathway ,RNA processing ,protein sf2 ,Ubiquitin-Conjugating Enzymes ,biology.protein ,RNA ,Posttranslational modification ,molecular recognition ,Tumor Suppressor Protein p53 ,metabolism ,SUMO 1 protein ,Heat-Shock Response - Abstract
Protein modification by conjugation of small ubiquitin-related modifier (SUMO) is involved in diverse biological functions, such as transcription regulation, subcellular partitioning, stress response, DNA damage repair, and chromatin remodeling. Here, we show that the serine/arginine-rich protein SF2/ASF, a factor involved in splicing regulation and other RNA metabolism-related processes, is a regulator of the sumoylation pathway. The overexpression of this protein stimulates, but its knockdown inhibits SUMO conjugation. SF2/ASF interacts with Ubc9 and enhances sumoylation of specific substrates, sharing characteristics with already described SUMO E3 ligases. In addition, SF2/ASF interacts with the SUMO E3 ligase PIAS1 (protein inhibitor of activated STAT-1), regulating PIAS1-induced overall protein sumoylation. The RNA recognition motif 2 of SF2/ASF is necessary and sufficient for sumoylation enhancement. Moreover, SF2/ASF has a role in heat shock-induced sumoylation and promotes SUMO conjugation to RNA processing factors. These results add a component to the sumoylation pathway and a previously unexplored role for the multifunctional SR protein SF2/ASF. Fil:Pelisch, F. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Gerez, J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Druker, J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Schor, I.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Muñoz, M.J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Risso, G. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Petrillo, E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Srebrow, A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
- Published
- 2010
31. Cytokines and genes in pituitary tumorigenesis: RSUME role in cell biology
- Author
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Mariana, Fuertes, Juan, Gerez, Mariana, Haedo, Damiana, Giacomini, Marcelo, Páez-Pereda, Marta, Labeur, Günter K, Stalla, and Eduardo, Arzt
- Subjects
Interleukin-6 ,Gene Expression Profiling ,Cytokine Receptor gp130 ,Animals ,Humans ,Pituitary Neoplasms ,Transcription Factors - Abstract
Cytokines of the IL-6 or gp130 family regulate many cellular responses and play regulatory roles in numerous tissues, and are placed as auto-paracrine regulators of pituitary function acting in normal and tumoral anterior pituitary cells. Especially, IL-6 has a regulatory role in the hormone secretion and growth of the anterior pituitary and is involved in adenoma pathogenesis. Recently, IL-6 has been shown to mediate oncogene-induced senescence (OIS). IL-6 might participate in such a process in adenomas pituitary as well. From pituitary tumoral gp130 overexpressing cells, an unknown protein, RSUME, has been cloned. RSUME is induced by hypoxia in pituitary tumors and regulate pathways involved in angiogenic and tumorigenic processes (NF-kappaB/IkappaB and HIF-1alpha pathways). Thus, it could have an important role in the development of the pituitary tumors.
- Published
- 2010
32. Cytokines and Genes in Pituitary Tumorigenesis: RSUME Role in Cell Biology
- Author
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Mariana Haedo, Günter K. Stalla, Juan Gerez, Marcelo Paez-Pereda, Damiana Giacomini, Mariana Fuertes, Eduardo Arzt, and Marta Labeur
- Subjects
Gene expression profiling ,Biology ,Pituitary tumorigenesis ,Glycoprotein 130 ,Gene ,Function (biology) ,Cell biology - Abstract
Cytokines of the IL-6 or gp130 family regulate many cellular responses and play regulatory roles in numerous tissues, and are placed as auto-paracrine regulators of pituitary function acting in normal
- Published
- 2010
33. Pituitary action of cytokines: focus on BMP-4 and gp130 family
- Author
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Juan Gerez, Marcelo Paez-Pereda, Alberto Carbia Nagashima, Damiana Giacomini, Matias Acuña, Eduardo Arzt, Günter K. Stalla, Susana Silberstein, Marily Theodoropoulou, Marta Labeur, and Ulrich Renner
- Subjects
medicine.medical_specialty ,Pituitary gland ,Endocrinology, Diabetes and Metabolism ,medicine.medical_treatment ,Bone Morphogenetic Protein 4 ,Biology ,gp130 family ,TGF- family ,Models, Biological ,Cellular and Molecular Neuroscience ,Endocrinology ,Anterior pituitary ,Internal medicine ,medicine ,Cytokine Receptor gp130 ,Animals ,Humans ,Pituitary action of cytokines ,Endocrine and Autonomic Systems ,food and beverages ,Glycoprotein 130 ,Glandula endocrina ,Cytokine ,medicine.anatomical_structure ,Multigene Family ,Pituitary Gland ,Bone Morphogenetic Proteins ,Cytokines ,Endocrine gland ,Hormone - Abstract
The anterior pituitary can develop benign tumors of different sizes, classified as micro- and macroadenomas, frequently associated with high levels of hormone production, leading to different associated syndromes like Cushing's disease, acromegaly or prolactinomas. Much work has been done in order to understand the signaling pathways and the factors and hormones involved in the pituitary tumorigenic process. In recent years, much evidence has been collected and it is now well documented that cytokines of the gp130 family, such as interleukin-6, that use gp130 as a common signaling protein stimulate not only the proliferation but also the hormone secretion of pituitary cells. Experiments in vivo have shown that the overexpression of the gp130 receptor resulted in pituitary abnormal growth. Moreover, it has been recently described that bone morphogenetic protein-4 (BMP-4), a member of the TGF-beta family, has a stimulatory role on lactosomatotropic cells promoting the development of prolactinomas but it has an inhibitory action on the corticotropic lineage. This inhibitory action prevents Cushing's disease progression. Furthermore, BMP-4 mediates the antiproliferative action of retinoic acid in these cells. The present review highlights the most recent work about gp130 and TGF-beta cytokine families and their role in pituitary tumorigenesis Fil: Giacomini, Damiana Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentina Fil: Acuña, Matias Nicolas. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina Fil: Gerez, Juan Atilio. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina Fil: Carbia Nagashima, Alberto Nicolas. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina Fil: Silberstein, Susana. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina Fil: Paez Pereda, Marcelo. Max Planck Institute of Psychiatry; Alemania Fil: Labeur, Marta. Max Planck Institute of Psychiatry; Alemania Fil: Theodoropoulou, Marily. Max Planck Institute of Psychiatry; Alemania Fil: Renner, Ulrich. Max Planck Institute of Psychiatry; Alemania Fil: Stalla, Günter K.. Max Planck Institute of Psychiatry; Alemania Fil: Arzt, Eduardo Simon. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentina
- Published
- 2007
34. Bone morphogenetic protein-4 inhibits corticotroph tumor cells: Involvement in the retinoic acid inhibitory action
- Author
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Marily Theodoropoulou, Juan Gerez, Marcelo Paez-Pereda, Silvia Berner, Marco Losa, Michael Buchfelder, Eduardo Arzt, Damiana Giacomini, Günter K. Stalla, Ulrich Renner, Damian Refojo, Marta Labeur, A. Chervin, Giacomini, Damiana, Páez-Pereda, Marcelo, Theodoropoulou, Marily, Labeur, Marta, Refojo, Damian, Gerez, Juan, Chervin, Alberto, Berner, Silvia, Losa, Marco, Buchfelder, Michael, Renner, Ulrich, Stalla, Günter K., and Arzt, Eduardo
- Subjects
cancer inhibition ,Retinoic acid ,Bone Morphogenetic Protein 4 ,animal cell ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,Endocrinology ,Reference Values ,TUMOR CELLS ,noggin ,retinoic acid ,carcinogenicity ,Reference Value ,Pituitary Neoplasm ,enzyme inhibition ,Cushing Syndrome ,comparative study ,transcription initiation ,0303 health sciences ,Smad4 protein ,Cushing disease ,disease course ,article ,cell line ,Immunohistochemistry ,3. Good health ,Bone morphogenetic protein 4 ,priority journal ,Pituitary Gland ,embryonic structures ,Bone Morphogenetic Proteins ,signal transduction ,Cell Division ,Adenoma ,medicine.medical_specialty ,animal structures ,animal experiment ,bone morphogenetic protein 4 ,030209 endocrinology & metabolism ,Tumor cells ,Tretinoin ,Biology ,animal tissue ,03 medical and health sciences ,Internal medicine ,Cell Line, Tumor ,medicine ,Animals ,Humans ,controlled study ,Pituitary Neoplasms ,human ,protein expression ,mouse ,030304 developmental biology ,Mice nude ,nonhuman ,Animal ,Bone Morphogenetic Protein ,animal model ,corticotropin release ,human tissue ,cell proliferation ,chemistry ,corticotropin ,genetic transfection ,adenohypophysis - Abstract
The molecular mechanisms governing the pathogenesis of ACTH-secreting pituitary adenomas are still obscure. Furthermore, the pharmacological treatment of these tumors is limited. In this study, we report that bone morphogenetic protein-4 (BMP-4) is expressed in the corticotrophs of human normal adenohypophysis and its expression is reduced in corticotrophinomas obtained from Cushing's patients compared with the normal pituitary. BMP-4 treatment of AtT-20 mouse corticotrophinoma cells has an inhibitory effect on ACTH secretion and cell proliferation. AtT-20 cells stably transfected with a dominant-negative form of the BMP-4 signal cotransducer Smad-4 or the BMP-4 inhibitor noggin have increased tumorigenicity in nude mice, showing that BMP-4 has an inhibitory role on corticotroph tumorigenesis in vivo. Because the activation of the retinoic acid receptor has an inhibitory action on Cushing's disease progression, we analyzed the putative interaction of these two pathways. Indeed, retinoic acid induces both BMP-4 transcription and expression and its antiproliferative action is blocked in Smad-4dn- and noggin-transfected Att-20 cells that do not respond to BMP-4. Therefore, retinoic acid induces BMP-4, which participates in the antiproliferative effects of retinoic acid. This new mechanism is a potential target for therapeutic approaches for Cushing's disease. Fil: Giacomini, Damiana Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina Fil: Páez Pereda, Marcelo. Max Planck Institute of Psychiatry; Alemania. Affectis Pharmaceuticals; Alemania Fil: Theodoropoulou, Marily. Max Planck Institute of Psychiatry; Alemania Fil: Labeur, Marta. Max Planck Institute of Psychiatry; Alemania Fil: Refojo, Damian. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina Fil: Gerez, Juan Atilio. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina Fil: Chervin, Alberto. Hospital Santa Lucía; Argentina Fil: Berner, Silvia. Hospital Santa Lucía; Argentina Fil: Losa, Marco. Max Planck Institute of Psychiatry; Alemania Fil: Buchfelder, Michael. University of Gottingen Medical School; Alemania Fil: Renner, Ulrich. Max Planck Institute of Psychiatry; Alemania Fil: Stalla, Günter K.. Max Planck Institute of Psychiatry; Alemania Fil: Arzt, Eduardo Simon. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina
- Published
- 2006
35. Bone Morphogenetic Protein-4 Control of Pituitary Pathophysiology
- Author
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A. Chervin, Damiana Giacomini, Silvia Berner, Marta Labeur, Ulrich Renner, Günter K. Stalla, Juan Gerez, Marcelo Paez-Pereda, Eduardo Arzt, Alberto Carbia Nagashima, Marily Theodoropoulou, and Damian Refojo
- Subjects
medicine.medical_specialty ,Bone morphogenetic protein 8A ,Biology ,Bone morphogenetic protein ,Bone morphogenetic protein 2 ,Bone morphogenetic protein 7 ,Bone morphogenetic protein 6 ,Bone morphogenetic protein 5 ,Endocrinology ,Bone morphogenetic protein 4 ,Internal medicine ,GDF10 ,Cancer research ,medicine - Abstract
Bone morphogenetic protein-4 (BMP-4), a member of the transforming growth factor-Beta(TGF-Beta) family, is overexpressed in different prolactinoma models and induces the development of these lineage adenomas. SMAD proteins activated by growth factors of the TGF-Beta and BMP family interact with estrogen receptors to stimulate the proliferation of prolactin and growth hormone-secreting cells. Furthermore, BMP-4 presents differential expression in normal and adenomatous corticotropes and inhibitory action on corticotropinoma cell proliferation. Moreover, BMP-4 mediates the antiproliferative action of retinoic acid in these cells. The present review highlights not only the crucial and opposite role of BMP-4 in the progression of pituitary adenomas but also that BMP-4 and retinoic acid interaction might serve as a potential new mechanism target for therapeutic approaches for Cushing disease.
- Published
- 2006
36. Subject Index Vol. 71, Suppl. 2, 2009
- Author
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Eivind Carlsen, Michelle T. Fleming, Karen P. Steel, Lars Sävendahl, Damiana Giacomini, Gavin Kelsey, Joel N. Hirschhorn, David B. Dunger, Guillaume Lettre, Ivo J.P. Arnhold, Eduardo Arzt, James W. MacDonald, Thomas Eggermann, Amanda H. Mortensen, Shannon W. Davis, Günter K. Stalla, Noboru Egashira, Catherine Molinas, Piero Carninci, Yoshihide Hayashizaki, Sunita K. Agarwal, Márta Korbonits, Stephen J. Marx, Wietske A. Ester, Carmen M. Mateo, Atsushi Ozawa, Debashis Ghosh, Linda Fryklund, Albert Beckers, Anita Hokken Koelega, Juan Gerez, Marcelo Paez-Pereda, Sally A. Camper, Vera Popovic-Brkic, Robert H. Lyons, Mehul T. Dattani, Marta Labeur, Andrea Lania, Andrei S. Chagin, Ron G. Rosenfeld, Mariana Haedo, Shlomo Melmed, Michelle L. Brinkmeier, Jimena Druker, Anna Spada, Maithé Tauber, Janina Caliebe, Alexander A. L. Jorge, Adrian J. L. Clark, L.B. Johnston, Wolter J. Mooi, Vera Chesnokova, Mary Anne Potok, Robert Yoshiyuki Osamura, Adrian F. Daly, Vivian Hwa, Maria A. Tichomirowa, Michael B. Ranke, Martin O. Savage, Gerhard Binder, and Hartmut A. Wollmann
- Subjects
Endocrinology ,Index (economics) ,Endocrinology, Diabetes and Metabolism ,Pediatrics, Perinatology and Child Health ,Statistics ,Subject (documents) ,Psychology - Published
- 2009
37. Non-targeted metabolomic approach reveals two distinct types of metabolic responses to telomerase dysfunction in S. cerevisiae
- Author
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Juan Gerez, Claus M. Azzalin, Madina Mansurova, Kyle A. Jay, Florian Buettner, Harry Wischnewski, Konstantins Jefimovs, Shady Saad, Alfredo J. Ibáñez, Reinhard Dechant, and Thomas Stadelmann
- Subjects
0301 basic medicine ,Genetics ,Telomerase ,DNA damage ,ved/biology ,Endocrinology, Diabetes and Metabolism ,Clinical Biochemistry ,ved/biology.organism_classification_rank.species ,Computational biology ,Biology ,Biochemistry ,Phenotype ,Yeast ,Telomere ,03 medical and health sciences ,030104 developmental biology ,Metabolomics ,Metabolome ,Model organism - Abstract
The alternative lengthening of telomeres (ALT) mechanism was first observed in the model organism S. cerevisiae. Interestingly, this mechanism is necessary for the viability of some tumor cells. Unfortunately, its molecular underpinnings are not yet completely understood. Here, we combine carefully designed non-targeted mass spectrometry-based metabolomics experiments with a bioinformatics approach to characterize the ALT positive phenotype observed in yeast at the metabolomics level. We profiled the metabolome using mass spectrometry in yeast strains that have lost telomerase expression, as well as that in pre-senescence and the rescued states. To dissect unwanted technical variation from biologically relevant variation between these states, we used a two-step normalization strategy, i.e., first, an empirical Bayesian framework; and next, we corrected for second-order technical effects. Our results show that ALT-positive yeast strains present two different types of metabolic responses to the genetically-induced telomerase dysfunction: (i) systemic and (ii) specific. The key-difference between these responses is that the systemic response lasts even after the yeast strains have been genetically rescued, while the specific response does not. Interestingly, these metabolic changes can be associated with generic stress responses (e.g., DNA damage) as well as specific responses like accelerated aging of early telomerase-inactivation. A mass spectrometry-based metabolomics approach reveals two distinct types of metabolomics response to telomerase dysfunction in yeast. By identifying these changes in protein (e.g., ARG7, and ARG1), and metabolite (e.g., dATP, and dDTP) amounts, we complement the available information on ALT at the genome-wide level.
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