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6 results on '"Lorène Belval"'

1. Structural basis of nanobody recognition of grapevine fanleaf virus and of virus resistance loss

Author

Olivier Lemaire, Christophe Ritzenthaler, Aurélie Marmonier, Sophie Gersch, Bruno P. Klaholz, Léa Ackerer, Corinne Schmitt-Keichinger, Vianney Poignavent, Kamal Hleibieh, Lorène Belval, Serge Muyldermans, Véronique Komar, Caroline Hemmer, Patrick Bron, Ahmed Ghannam, Claude Sauter, Igor Orlov, Pascale Schellenberger, Jean-Michel Hily, Gérard Demangeat, Bernard Lorber, Emmanuelle Vigne, Klaholz, Bruno, Appel à projets générique - Résistances antivirales combinées pour lutter contre la maladie du court-noué de la vigne - - COMBiNiNG2014 - ANR-14-CE19-0022 - Appel à projets générique - VALID, Infrastructure Française pour la Biologie Structurale Intégrée - - FRISBI2010 - ANR-10-INBS-0005 - INBS - VALID, Centre for Integrative Biology - CBI (Inserm U964 - CNRS UMR7104 - IGBMC), 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), Institut de biologie moléculaire des plantes (IBMP), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Santé de la vigne et qualité du vin (SVQV), Université de Strasbourg (UNISTRA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Architecture et Réactivité de l'ARN (ARN), Institut de biologie moléculaire et cellulaire (IBMC), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut Français de la Vigne et du Vin [Le Grau du Roi], Centre de Biochimie Structurale [Montpellier] (CBS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Vrije Universiteit Brussel (VUB), ANR-14-CE19-0022,COMBiNiNG,Résistances antivirales combinées pour lutter contre la maladie du court-noué de la vigne(2014), ANR-10-INBS-0005,FRISBI,Infrastructure Française pour la Biologie Structurale Intégrée(2010), Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), and Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
0106 biological sciences, Models, Molecular, GFLV, Nematoda, Cryo-electron microscopy, Protein Conformation, [SDV]Life Sciences [q-bio], Nepovirus, Biology, Antibodies, Viral, 01 natural sciences, Virus, Epitope, Plant Viruses, 03 medical and health sciences, Epitopes, Capsid, Plant virus, structural biology, Animals, Vitis, Binding site, 030304 developmental biology, Plant Diseases, 0303 health sciences, Multidisciplinary, Cryoelectron Microscopy, Grapevine fanleaf virus, virus, Biological Sciences, biology.organism_classification, Virology, [SDV] Life Sciences [q-bio], Plant Leaves, nanobody, Structural biology, Capsid Proteins, 010606 plant biology & botany, Single-Chain Antibodies
Abstract

International audience; Grapevine fanleaf virus (GFLV) is a picorna-like plant virus transmitted by nematodes that affects vineyards worldwide. Nanobody (Nb)-mediated resistance against GFLV has been created recently, and shown to be highly effective in plants, including grapevine, but the underlying mechanism is unknown. Here we present the high-resolution cryo electron microscopy structure of the GFLV-Nb23 complex, which provides the basis for molecular recognition by the Nb. The structure reveals a composite binding site bridging over three domains of one capsid protein (CP) monomer. The structure provides a precise mapping of the Nb23 epitope on the GFLV capsid in which the antigen loop is accommodated through an induced-fit mechanism. Moreover, we uncover and characterize several resistance-breaking GFLV isolates with amino acids mapping within this epitope, including C-terminal extensions of the CP, which would sterically interfere with Nb binding. Escape variants with such extended CP fail to be transmitted by nematodes linking Nb-mediated resistance to vector transmission. Together, these data provide insights into the molecular mechanism of Nb23-mediated recognition of GFLV and of virus resistance loss.

Published
2020
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2. From a Movement-Deficient Grapevine Fanleaf Virus to the Identification of a New Viral Determinant of Nematode Transmission

Author

Lorène Belval, Corinne Schmitt-Keichinger, Christophe Ritzenthaler, P. Andret-Link, Sophie Gersch, Aurélie Marmonier, Véronique Komar, Olivier Lemaire, Gérard Demangeat, Emmanuelle Vigne, Santé de la vigne et qualité du vin (SVQV), Institut National de la Recherche Agronomique (INRA)-Université de Strasbourg (UNISTRA), Institut de biologie moléculaire des plantes (IBMP), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), and Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)

Subjects
Models, Molecular, 0106 biological sciences, Nematoda, Protein Conformation, nematode, viruses, Nepovirus, Xiphinema, Disease Vectors, 3D structure, 01 natural sciences, Article, Virus, Arabis mosaic virus, Structure-Activity Relationship, Viral Proteins, 03 medical and health sciences, Genes, Reporter, Virology, Plant virus, capsid, Animals, [SDV.BV]Life Sciences [q-bio]/Vegetal Biology, Amino Acid Sequence, Plant Diseases, 030304 developmental biology, Recombination, Genetic, 0303 health sciences, biology, viral determinant, transmission, Grapevine fanleaf virus, biology.organism_classification, grapevine, Infectious Diseases, Nematode, Capsid, RNA, Viral, movement, tubule, 010606 plant biology & botany
Abstract

Grapevine fanleaf virus (GFLV) and arabis mosaic virus (ArMV) are nepoviruses responsible for grapevine degeneration. They are specifically transmitted from grapevine to grapevine by two distinct ectoparasitic dagger nematodes of the genus Xiphinema. GFLV and ArMV move from cell to cell as virions through tubules formed into plasmodesmata by the self-assembly of the viral movement protein. Five surface-exposed regions in the coat protein called R1 to R5, which differ between the two viruses, were previously defined and exchanged to test their involvement in virus transmission, leading to the identification of region R2 as a transmission determinant. Region R4 (amino acids 258 to 264) could not be tested in transmission due to its requirement for plant systemic infection. Here, we present a fine-tuning mutagenesis of the GFLV coat protein in and around region R4 that restored the virus movement and allowed its evaluation in transmission. We show that residues T258, M260, D261, and R301 play a crucial role in virus transmission, thus representing a new viral determinant of nematode transmission.

Published
2019
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3. Nanobody-mediated resistance to Grapevine fanleaf virus in plants

Author

Kamal Hleibieh, Lorène Belval, Serge Muyldermans, François Berthold, Sophie Gersch, Carlos Gutierrez, Claude Gertz, Gérard Demangeat, Samia Djennane, Véronique Komar, Aurélie Marmonier, Mireille Perrin, Corinne Schmitt-Keichinger, Emmanuelle Vigne, Christophe Ritzenthaler, Baptiste Monsion, Olivier Lemaire, Léa Ackerer, Bernard Lorber, Shahinez Garcia, Caroline Hemmer, Institut de biologie moléculaire des plantes (IBMP), Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), Architecture et réactivité de l'ARN (ARN), Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), Architecture et Réactivité de l'ARN (ARN), Institut de biologie moléculaire et cellulaire (IBMC), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique ( INRA) 'Plant Health and the Environment' division, Agence Nationale de la Recherche (ANR) award COMBiNiNG [ANR-14-CE19-0022], Agence Nationale de la Recherche (ANR) award VinoBodies [ANR-14-CE19-0018], European Regional Development Fund (ERDF), Conseil interprofessionel des vins d'Alsace, Comite interprofessionel du vin Champagne, Bureau interprofessionnel des vins de Bourgogne, Conseil interprofessionnel du vin de Bordeaux, Region Alsace, CIFRE grant from the Institut Francais de la Vigne et du Vin - ANRT [2012/0929], ANR-14-CE19-0022,COMBiNiNG,Résistances antivirales combinées pour lutter contre la maladie du court-noué de la vigne(2014), ANR-14-CE19-0018,VinoBodies,Nanobodies: le couteau Suisse de la virologie de la vigne(2014), Santé de la vigne et qualité du vin (SVQV), Institut National de la Recherche Agronomique (INRA)-Université de Strasbourg (UNISTRA), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Demangeat, Gérard, Department of Bio-engineering Sciences, and Cellular and Molecular Immunology

Subjects
0106 biological sciences, 0301 basic medicine, single-chain antibodies, viruses, Nepovirus, Nicotiana benthamiana, Plant Science, 01 natural sciences, Virus, Plant Viruses, Arabis mosaic virus, 03 medical and health sciences, plant virus, Viral life cycle, Plant virus, Plant Viruses/genetics, [SDV.BV]Life Sciences [q-bio]/Vegetal Biology, Plant Diseases/immunology, GMO, nanobodies, transgenic plant, grapevine, nepovirus, Research Articles, Plant Diseases, 2. Zero hunger, Single-Domain Antibodies/genetics, single‐chain antibodies, biology, Host (biology), food and beverages, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Grapevine fanleaf virus, Single-Domain Antibodies, biology.organism_classification, Virology, Nepovirus/pathogenicity, 3. Good health, 030104 developmental biology, Agronomy and Crop Science, Research Article, 010606 plant biology & botany, Biotechnology
Abstract

International audience; Since their discovery, single-domain antigen-binding fragments of camelid-derived heavy-chainonly antibodies, also known as nanobodies (Nbs), have proven to be of outstanding interest as therapeutics against human diseases and pathogens including viruses, but their use against phytopathogens remains limited. Many plant viruses including Grapevine fanleaf virus (GFLV), a nematode-transmitted icosahedral virus and causal agent of fanleaf degenerative disease, have worldwide distribution and huge burden on crop yields representing billions of US dollars of losses annually, yet solutions to combat these viruses are often limited or inefficient. Here, we identified a Nb specific to GFLV that confers strong resistance to GFLV upon stable expression in the model plant Nicotiana benthamiana and also in grapevine rootstock, the natural host of the virus. We showed that resistance was effective against a broad range of GFLV isolates independently of the inoculation method including upon nematode transmission but not againstits close relative, Arabis mosaic virus. We also demonstrated that virus neutralization occurs at an early step of the virus life cycle, prior to cell-to-cell movement. Our findings will not only be instrumental to confer resistance to GFLV in grapevine, but more generally they pave the way for the generation of novel antiviral strategies in plants based on Nbs

Published
2017
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4. Structural basis of nanobody-recognition of grapevine fanleaf virus and of virus resistance loss

Author

Bernard Lorber, Sophie Gersch, Emmanuelle Vigne, Corinne Schmitt-Keichinger, Serge Muyldermans, Kamal Hleibieh, Vianney Poignavent, Lorène Belval, Patrick Bron, Pascale Schellenberger, Jean-Michel Hily, Gérard Demangeat, Bruno P. Klaholz, Ahmed Ghannam, Léa Ackerer, Igor Orlov, Véronique Komar, Olivier Lemaire, Caroline Hemmer, Christophe Ritzenthaler, Aurélie Marmonier, Claude Sauter, Institut de génétique et biologie moléculaire et cellulaire (IGBMC), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Louis Pasteur - Strasbourg I, 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), Institut de biologie moléculaire des plantes (IBMP), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Architecture et réactivité de l'ARN (ARN), Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), Architecture et Réactivité de l'ARN (ARN), Institut de biologie moléculaire et cellulaire (IBMC), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), ANR-14-CE19-0022,COMBiNiNG,Résistances antivirales combinées pour lutter contre la maladie du court-noué de la vigne(2014), ANR-10-INBS-0005,FRISBI,Infrastructure Française pour la Biologie Structurale Intégrée(2010), Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)

Subjects
0106 biological sciences, Genetics, 0303 health sciences, biology, Grapevine fanleaf virus, Virus resistance, biology.organism_classification, 01 natural sciences, Epitope, 03 medical and health sciences, Molecular recognition, Capsid, Plant virus, [SDV.BV]Life Sciences [q-bio]/Vegetal Biology, Binding site, 030304 developmental biology, 010606 plant biology & botany, Conformational epitope
Abstract

Grapevine fanleaf virus (GFLV) is a picorna-like plant virus transmitted by nematodes that affects vineyards worldwide. Nanobody (Nb)-mediated resistance against GFLV has been created recently and shown to be highly effective in plants including grapevine, but the underlying mechanism is unknown. Here we present the high-resolution cryo-EM structure of the GFLV-Nb23 complex which provides the basis for the molecular recognition by the nanobody. The structure reveals a composite binding site bridging over 3 domains of the capsid protein (CP) monomer. The structure provides a precise mapping of the Nb23 epitope on the GFLV capsid in which the antigen loop is accommodated through an induced fit mechanism. Moreover, we uncover and characterize several resistance-breaking GFLV isolates with amino acids mapping within this epitope, including C-terminal extensions of the CP, which would sterically interfere with Nb binding. Escape variants with such extended CP fail to be transmitted by nematodes linking Nb-mediated resistance to vector transmission. Together, these data provide insights into the molecular mechanism of Nb23-mediated recognition of GFLV and of virus resistance loss.SignificanceGrapevine fanleaf virus (GFLV) is a picorna-like plant virus that severely impacts vineyards worldwide. While Nanobodies (Nb) confer resistance to GFLV in plants the underlying molecular mechanism of action is unknown. Here we present the high-resolution cryo-EM structure of the GFLV-Nb complex. It uncovers the conformational epitope on the capsid surface which is a composite binding site into which the antigen loop is accommodated through an induced fit mechanism. Furthermore, we describe several resistance-breaking isolates of GFLV with reduced Nb binding capacity. Those that carry a C-terminal extension also fail to be transmitted by nematodes. Together, these data provide structure-function insights into the Nb-GFLV recognition and the molecular mechanism leading to loss of resistance.

Published
2019
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5. Ectoparasitic Nematode Vectors of Grapevine Viruses

Author

Gérard Demangeat, P. Andret-Link, Christophe Ritzenthaler, Aurélie Marmonier, Kamal Hleibieh, Lorène Belval, ProdInra, Migration, Springer, Santé de la vigne et qualité du vin (SVQV), Institut National de la Recherche Agronomique (INRA)-Université de Strasbourg (UNISTRA), Institut de biologie moléculaire des plantes (IBMP), Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), and Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0106 biological sciences, 0301 basic medicine, [SDE] Environmental Sciences, [SDV]Life Sciences [q-bio], Nepovirus, 01 natural sciences, Virus, 03 medical and health sciences, Xiphinema, [SDV.BV]Life Sciences [q-bio]/Vegetal Biology, Transmission, [SDV.BV] Life Sciences [q-bio]/Vegetal Biology, Vector (molecular biology), Fanleaf degeneration, Longidorus, Longidoridae, Genetics, Rhizosphere, biology, biology.organism_classification, Virology, [SDV] Life Sciences [q-bio], 030104 developmental biology, Nematode, [SDE]Environmental Sciences, Specificity, Grapevine, Vector, 010606 plant biology & botany
Abstract

International audience; Nematodes of the genera Longidorus, Paralongidorus, and Xiphinema in the family Longidoridae can parasitize grapevine roots. These ectoparasitic soil-borne nematodes live in proximity to the rhizosphere and use a stylet to feed on root cells. During the feeding process, some nematodes are able to acquire and transmit nepoviruses, the causal agents of grapevine degeneration disease. The association between nematode vectors and the virus(es) they transmit is specific. This specificity is linked to the nature, site, and mechanism of virus retention. Here, we present and discuss major features of the interaction between nematodes and viruses with regard to the biology, ecology, and life cycle of the nematodes. In addition, we discuss how reverse genetics in combination with virus structural data have recently provided insights into the specificity of the transmission process through the identification of a cavity-like domain on the virion surface that could be recognized by a ligand within the nematode feeding apparatus. Finally, we offer some perspectives on future research to further advance nematode-virus interactions.

Published
2017
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6. A strain-specific segment of the RNA-dependent RNA polymerase of grapevine fanleaf virus determines symptoms in Nicotiana species

Author

Véronique Komar, Léa Ackerer, Emmanuelle Vigne, John Gottula, Olivier Lemaire, Christophe Ritzenthaler, Lalaina Rakotomalala, Corinne Schmitt-Keichinger, Lorène Belval, Marc Fuchs, Institut de biologie moléculaire des plantes (IBMP), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Laboratoire SUBATECH Nantes (SUBATECH), Mines Nantes (Mines Nantes)-Université de Nantes (UN)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), SLAC National Accelerator Laboratory (SLAC), Stanford University, Santé de la vigne et qualité du vin (SVQV), Institut National de la Recherche Agronomique (INRA)-Université Louis Pasteur - Strasbourg I, Department of Plant Pathology and Plant-Microbe Biology, Cornell University, Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA), INRA, Department Sante des Plantes et Environnement of INRA, Hatch Act Formula and Nolan Funds, and USDA-NIFA

Subjects
0106 biological sciences, [SDV]Life Sciences [q-bio], viruses, Molecular Sequence Data, Nepovirus, RNA-dependent RNA polymerase, Nicotiana benthamiana, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, 01 natural sciences, Virus, Viral Proteins, 03 medical and health sciences, chemistry.chemical_compound, Species Specificity, Virology, RNA polymerase, Tobacco, Secoviridae, [SDV.BV]Life Sciences [q-bio]/Vegetal Biology, Vitis, Amino Acid Sequence, Phylogeny, ComputingMilieux_MISCELLANEOUS, Plant Diseases, 030304 developmental biology, 0303 health sciences, biology, fungi, food and beverages, RNA, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Grapevine fanleaf virus, Sequence Analysis, DNA, RNA-Dependent RNA Polymerase, biology.organism_classification, Molecular biology, 3. Good health, chemistry, RNA, Viral, 010606 plant biology & botany
Abstract

Factors involved in symptom expression of viruses from the genus Nepovirus in the family Secoviridae such as grapevine fanleaf virus (GFLV) are poorly characterized. To identify symptom determinants encoded by GFLV, infectious cDNA clones of RNA1 and RNA2 of strain GHu were developed and used alongside existing infectious cDNA clones of strain F13 in a reverse genetics approach. In vitro transcripts of homologous combinations of RNA1 and RNA2 induced systemic infection in Nicotiana benthamiana and Nicotiana clevelandii with identical phenotypes to WT virus strains, i.e. vein clearing and chlorotic spots on N. benthamiana and N. clevelandii for GHu, respectively, and lack of symptoms on both hosts for F13. The use of assorted transcripts mapped symptom determinants on RNA1 of GFLV strain GHu, in particular within the distal 408 nt of the RNA-dependent RNA polymerase (1EPol), as shown by RNA1 transcripts for which coding regions or fragments derived thereof were swapped. Semi-quantitative analyses indicated no significant differences in virus titre between symptomatic and asymptomatic plants infected with various recombinants. Also, unlike the nepovirus tomato ringspot virus, no apparent proteolytic cleavage of GFLV protein 1EPol was detected upon virus infection or transient expression in N. benthamiana. In addition, GFLV protein 1EPol failed to suppress silencing of EGFP in transgenic N. benthamiana expressing EGFP or to enhance GFP expression in patch assays in WT N. benthamiana. Together, our results suggest the existence of strain-specific functional domains, including a symptom determinant module, on the RNA-dependent RNA polymerase of GFLV.

Published
2013
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