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Accueil > Départements > Virologie > Yves GAUDIN : Rhabdovirus

Publications de l’équipe



  • E. Baquero, A. A. Albertini, H. Raux, A. Abou‐Hamdan, E. Boeri‐Erba, M. Ouldali, L. Buonocore, J. K. Rose, J. Lepault, S. Bressanelli, et Y. Gaudin, « Structural intermediates in the fusion‐associated transition of vesiculovirus glycoprotein », The EMBO Journal, vol. 36, nᵒ 5, p. 679-692, mars 2017.
    Mots-clés : B3S, conformational change, glycoprotein, IMAPP, intermediate structures, membrane fusion, RHABDO, Vesiculovirus, VIRO, VIROEM.

  • J. Nikolic et D. Blondel, « [Rabies virus induces the formation of stress granules, which are in close proximity to viral factories] », Medecine Sciences: M/S, vol. 33, nᵒ 11, p. 921-923, nov. 2017.

  • J. Nikolic, R. Le Bars, Z. Lama, N. Scrima, C. Lagaudrière-Gesbert, Y. Gaudin, et D. Blondel, « Negri bodies are viral factories with properties of liquid organelles », Nature Communications, vol. 8, nᵒ 1, p. 58, juill. 2017.
    Résumé : Replication of Mononegavirales occurs in viral factories which form inclusions in the host-cell cytoplasm. For rabies virus, those inclusions are called Negri bodies (NBs). We report that NBs have characteristics similar to those of liquid organelles: they are spherical, they fuse to form larger structures, and they disappear upon hypotonic shock. Their liquid phase is confirmed by FRAP experiments. Live-cell imaging indicates that viral nucleocapsids are ejected from NBs and transported along microtubules to form either new virions or secondary viral factories. Coexpression of rabies virus N and P proteins results in cytoplasmic inclusions recapitulating NBs properties. This minimal system reveals that an intrinsically disordered domain and the dimerization domain of P are essential for Negri bodies-like structures formation. We suggest that formation of liquid viral factories by phase separation is common among Mononegavirales and allows specific recruitment and concentration of viral proteins but also the escape to cellular antiviral response.Negative strand RNA viruses, such as rabies virus, induce formation of cytoplasmic inclusions for genome replication. Here, Nikolic et al. show that these so-called Negri bodies (NBs) have characteristics of liquid organelles and they identify the minimal protein domains required for NB formation.
    Mots-clés : IMAGIF, PF, PHOT, RHABDO, VIRO.


  • A. Brice, D. R. Whelan, N. Ito, K. Shimizu, L. Wiltzer-Bach, C. Y. Lo, D. Blondel, D. A. Jans, T. D. M. Bell, et G. W. Moseley, « Quantitative Analysis of the Microtubule Interaction of Rabies Virus P3 Protein: Roles in Immune Evasion and Pathogenesis », Scientific Reports, vol. 6, p. 33493, sept. 2016.
    Résumé : Although microtubules (MTs) are known to have important roles in intracellular transport of many viruses, a number of reports suggest that specific viral MT-associated proteins (MAPs) target MTs to subvert distinct MT-dependent cellular processes. The precise functional importance of these interactions and their roles in pathogenesis, however, remain largely unresolved. To assess the association with disease of the rabies virus (RABV) MAP, P3, we quantitatively compared the phenotypes of P3 from a pathogenic RABV strain, Nishigahara (Ni) and a non-pathogenic Ni-derivative strain, Ni-CE. Using confocal/live-cell imaging and dSTORM super-resolution microscopy to quantify protein interactions with the MT network and with individual MT filaments, we found that the interaction by Ni-CE-P3 is significantly impaired compared with Ni-P3. This correlated with an impaired capacity to effect association of the transcription factor STAT1 with MTs and to antagonize interferon (IFN)/STAT1-dependent antiviral signaling. Importantly, we identified a single mutation in Ni-CE-P3 that is sufficient to inhibit MT-association and IFN-antagonist function of Ni-P3, and showed that this mutation alone attenuates the pathogenicity of RABV. These data provide evidence that the viral protein-MT interface has important roles in pathogenesis, suggesting that this interface could provide targets for vaccine/antiviral drug development.
    Mots-clés : RHABDO, VIRO.

  • D. Liger, L. Mora, N. Lazar, S. Figaro, J. Henri, N. Scrima, R. H. Buckingham, H. van Tilbeurgh, V. Heurgué-Hamard, et M. Graille, « Mechanism of activation of methyltransferases involved in translation by the Trm112 ‘hub’ protein », Nucleic Acids Research, vol. 44, nᵒ 3, p. 1482-1482, févr. 2016.
    Mots-clés : B3S, FAAM, RHABDO, VIRO.

  • G. Maarifi, Z. Hannoun, M. C. Geoffroy, F. El Asmi, K. Zarrouk, S. Nisole, D. Blondel, et M. K. Chelbi-Alix, « MxA Mediates SUMO-Induced Resistance to Vesicular Stomatitis Virus », Journal of Virology, vol. 90, nᵒ 14, p. 6598-6610, juill. 2016.
    Résumé : Multiple cellular pathways are regulated by small ubiquitin-like modifier (SUMO) modification, including ubiquitin-mediated proteolysis, signal transduction, innate immunity, and antiviral defense. In the study described in this report, we investigated the effects of SUMO on the replication of two members of the Rhabdoviridae family, vesicular stomatitis virus (VSV) and rabies virus (RABV). We show that stable expression of SUMO in human cells confers resistance to VSV infection in an interferon-independent manner. We demonstrate that SUMO expression did not alter VSV entry but blocked primary mRNA synthesis, leading to a reduction of viral protein synthesis and viral production, thus protecting cells from VSV-induced cell lysis. MxA is known to inhibit VSV primary transcription. Interestingly, we found that the MxA protein was highly stabilized in SUMO-expressing cells. Furthermore, extracts from cells stably expressing SUMO exhibited an increase in MxA oligomers, suggesting that SUMO plays a role in protecting MxA from degradation, thus providing a stable intracellular pool of MxA available to combat invading viruses. Importantly, MxA depletion in SUMO-expressing cells abrogated the anti-VSV effect of SUMO. Furthermore, SUMO expression resulted in interferon-regulatory factor 3 (IRF3) SUMOylation, subsequently decreasing RABV-induced IRF3 phosphorylation and interferon synthesis. As expected, this rendered SUMO-expressing cells more sensitive to RABV infection, even though MxA was stabilized in SUMO-expressing cells, since its expression did not confer resistance to RABV. Our findings demonstrate opposing effects of SUMO expression on two viruses of the same family, intrinsically inhibiting VSV infection through MxA stabilization while enhancing RABV infection by decreasing IFN induction. IMPORTANCE: We report that SUMO expression reduces interferon synthesis upon RABV or VSV infection. Therefore, SUMO renders cells more sensitive to RABV but unexpectedly renders cells resistant to VSV by blocking primary mRNA synthesis. Unlike the interferon-mediated innate immune response, intrinsic antiviral resistance is mediated by constitutively expressed restriction factors. Among the various anti-VSV restriction factors, only MxA is known to inhibit VSV primary transcription, and we show here that its expression does not alter RABV infection. Interestingly, MxA depletion abolished the inhibition of VSV by SUMO, demonstrating that MxA mediates SUMO-induced intrinsic VSV resistance. Furthermore, MxA oligomerization is known to be critical for its protein stability, and we show that higher levels of oligomers were formed in cells expressing SUMO than in wild-type cells, suggesting that SUMO may play a role in protecting MxA from degradation, providing a stable intracellular pool of MxA able to protect cells from viral infection.
    Mots-clés : RHABDO, VIRO.

  • J. Nikolic, A. Civas, Z. Lama, C. Lagaudrière-Gesbert, et D. Blondel, « Rabies Virus Infection Induces the Formation of Stress Granules Closely Connected to the Viral Factories », PLoS pathogens, vol. 12, nᵒ 10, p. e1005942, oct. 2016.
    Résumé : Stress granules (SGs) are membrane-less dynamic structures consisting of mRNA and protein aggregates that form rapidly in response to a wide range of environmental cellular stresses and viral infections. They act as storage sites for translationally silenced mRNAs under stress conditions. During viral infection, SG formation results in the modulation of innate antiviral immune responses, and several viruses have the ability to either promote or prevent SG assembly. Here, we show that rabies virus (RABV) induces SG formation in infected cells, as revealed by the detection of SG-marker proteins Ras GTPase-activating protein-binding protein 1 (G3BP1), T-cell intracellular antigen 1 (TIA-1) and poly(A)-binding protein (PABP) in the RNA granules formed during viral infection. As shown by live cell imaging, RABV-induced SGs are highly dynamic structures that increase in number, grow in size by fusion events, and undergo assembly/disassembly cycles. Some SGs localize in close proximity to cytoplasmic viral factories, known as Negri bodies (NBs). Three dimensional reconstructions reveal that both structures remain distinct even when they are in close contact. In addition, viral mRNAs synthesized in NBs accumulate in the SGs during viral infection, revealing material exchange between both compartments. Although RABV-induced SG formation is not affected in MEFs lacking TIA-1, TIA-1 depletion promotes viral translation which results in an increase of viral replication indicating that TIA-1 has an antiviral effect. Inhibition of PKR expression significantly prevents RABV-SG formation and favors viral replication by increasing viral translation. This is correlated with a drastic inhibition of IFN-B gene expression indicating that SGs likely mediate an antiviral response which is however not sufficient to fully counteract RABV infection.
    Mots-clés : RHABDO, VIRO.


  • E. Baquero, A. A. V. Albertini, et Y. Gaudin, « Recent mechanistic and structural insights on class III viral fusion glycoproteins », Current Opinion in Structural Biology, vol. 33, p. 52-60, août 2015.
    Résumé : Enveloped viruses enter the cell by fusing their envelope with a cellular membrane. Fusion is catalyzed by conformational changes of viral glycoproteins from pre-fusion to post-fusion states. Structural studies have defined three classes of viral fusion glycoproteins. Class III comprises the fusion glycoproteins from rhabdoviruses (G), herpesviruses (gB), and baculoviruses (GP64). Although sharing the same fold, those glycoproteins exhibit striking differences in their modes of activation and interaction with the target membrane. Furthermore, for gB and GP64, only the post-fusion structure is known and the extent of their conformational change is still an unresolved issue. Further structural studies are therefore required to get a detailed insight in the working of those fusion machines.
    Mots-clés : Baculoviridae, Cell Membrane, Herpesviridae, Hydrogen-Ion Concentration, Protein Conformation, RHABDO, Rhabdoviridae, Viral Fusion Proteins, VIRO.

  • E. Baquero, A. A. Albertini, H. Raux, L. Buonocore, J. K. Rose, S. Bressanelli, et Y. Gaudin, « Structure of the Low pH Conformation of Chandipura Virus G Reveals Important Features in the Evolution of the Vesiculovirus Glycoprotein », PLOS Pathogens, vol. 11, nᵒ 3, p. e1004756, mars 2015.
    Mots-clés : B3S, Evolution, Molecular, Humans, Hydrogen-Ion Concentration, IMAPP, Nucleocapsid, Protein Structure, Tertiary, RHABDO, Vesiculovirus, Viral Fusion Proteins, VIRO.

  • F. Beilstein, L. Obiang, H. Raux, et Y. Gaudin, « Characterization of the Interaction between the Matrix Protein of Vesicular Stomatitis Virus and the Immunoproteasome Subunit LMP2 », Journal of Virology, vol. 89, nᵒ 21, p. 11019-11029, nov. 2015.
    Résumé : The matrix protein (M) of vesicular stomatitis virus (VSV) is involved in virus assembly, budding, gene regulation, and cellular pathogenesis. Using a yeast two-hybrid system, the M globular domain was shown to interact with LMP2, a catalytic subunit of the immunoproteasome (which replaces the standard proteasome catalytic subunit PSMB6). The interaction was validated by coimmunoprecipitation of M and LMP2 in VSV-infected cells. The sites of interaction were characterized. A single mutation of M (I96A) which significantly impairs the interaction between M and LMP2 was identified. We also show that M preferentially binds to the inactive precursor of LMP2 (bearing an N-terminal propeptide which is cleaved upon LMP2 maturation). Furthermore, taking advantage of a sequence alignment between LMP2 and its proteasome homolog, PSMB6 (which does not bind to M), we identified a mutation (L45R) in the S1 pocket where the protein substrate binds prior to cleavage and a second one (D17A) of a conserved residue essential for the catalytic activity, resulting in a reduction of the level of binding to M. The combination of both mutations abolishes the interaction. Taken together, our data indicate that M binds to LMP2 before its incorporation into the immunoproteasome. As the immunoproteasome promotes the generation of major histocompatibility complex (MHC) class I-compatible peptides, a feature which favors the recognition and the elimination of infected cells by CD8 T cells, we suggest that M, by interfering with the immunoproteasome assembly, has evolved a mechanism that allows infected cells to escape detection and elimination by the immune system. IMPORTANCE: The immunoproteasome promotes the generation of MHC class I-compatible peptides, a feature which favors the recognition and the elimination of infected cells by CD8 T cells. Here, we report on the association of vesicular stomatitis virus (VSV) matrix protein (M) with LMP2, one of the immunoproteasome-specific catalytic subunits. M preferentially binds to the LMP2 inactive precursor. The M-binding site on LMP2 is facing inwards in the immunoproteasome and is therefore not accessible to M after its assembly. Hence, M binds to LMP2 before its incorporation into the immunoproteasome. We suggest that VSV M, by interfering with the immunoproteasome assembly, has evolved a mechanism that allows infected cells to escape detection and elimination by the immune system. Modulating this M-induced immunoproteasome impairment might be relevant in order to optimize VSV for oncolytic virotherapy.
    Mots-clés : Base Sequence, Blotting, Western, Cysteine Endopeptidases, HeLa Cells, Humans, Immunoprecipitation, Molecular Sequence Data, Mutation, Protein Binding, RHABDO, Sequence Alignment, Sequence Analysis, DNA, Two-Hybrid System Techniques, Vesiculovirus, Viral Matrix Proteins, VIRO.

  • D. Blondel, G. Maarifi, S. Nisole, et M. K. Chelbi-Alix, « Resistance to Rhabdoviridae Infection and Subversion of Antiviral Responses », Viruses, vol. 7, nᵒ 7, p. 3675-3702, juill. 2015.
    Résumé : Interferon (IFN) treatment induces the expression of hundreds of IFN-stimulated genes (ISGs). However, only a selection of their products have been demonstrated to be responsible for the inhibition of rhabdovirus replication in cultured cells; and only a few have been shown to play a role in mediating the antiviral response in vivo using gene knockout mouse models. IFNs inhibit rhabdovirus replication at different stages via the induction of a variety of ISGs. This review will discuss how individual ISG products confer resistance to rhabdoviruses by blocking viral entry, degrading single stranded viral RNA, inhibiting viral translation or preventing release of virions from the cell. Furthermore, this review will highlight how these viruses counteract the host IFN system.
    Mots-clés : Animals, Humans, interferon, Interferons, ISG, rabies virus, RHABDO, Rhabdoviridae, Rhabdoviridae Infections, rhabdoviruses, vesicular stomatitis virus, VIRO.

  • B. Fouquet, J. Nikolic, F. Larrous, H. Bourhy, C. Wirblich, C. Lagaudrière-Gesbert, et D. Blondel, « Focal adhesion kinase is involved in rabies virus infection through its interaction with viral phosphoprotein P », Journal of Virology, vol. 89, nᵒ 3, p. 1640-1651, févr. 2015.
    Résumé : The rabies virus (RABV) phosphoprotein P is a multifunctional protein: it plays an essential role in viral transcription and replication, and in addition, RABV P has been identified as an interferon antagonist. Here, a yeast two-hybrid screen revealed that RABV P interacts with the focal adhesion kinase (FAK). The binding involved the 106-to-131 domain, corresponding to the dimerization domain of P and the C-terminal domain of FAK containing the proline-rich domains PRR2 and PRR3. The P-FAK interaction was confirmed in infected cells by coimmunoprecipitation and colocalization of FAK with P in Negri bodies. By alanine scanning, we identified a single mutation in the P protein that abolishes this interaction. The mutant virus containing a substitution of Ala for Arg in position 109 in P (P.R109A), which did not interact with FAK, is affected at a posttranscriptional step involving protein synthesis and viral RNA replication. Furthermore, FAK depletion inhibited viral protein expression in infected cells. This provides the first evidence of an interaction of RABV with FAK that positively regulates infection. IMPORTANCE: Rabies virus exhibits a small genome that encodes a limited number of viral proteins. To maintain efficient virus replication, some of them are multifunctional, such as the phosphoprotein P. We and others have shown that P establishes complex networks of interactions with host cell components. These interactions have revealed much about the role of P and about host-pathogen interactions in infected cells. Here, we identified another cellular partner of P, the focal adhesion kinase (FAK). Our data shed light on the implication of FAK in RABV infection and provide evidence that P-FAK interaction has a proviral function.
    Mots-clés : Animals, Cell Line, DNA Mutational Analysis, Focal Adhesion Protein-Tyrosine Kinases, Host-Pathogen Interactions, Humans, Immunoprecipitation, Inclusion Bodies, Viral, Microscopy, Confocal, Mutagenesis, Site-Directed, Phosphoproteins, Protein Binding, Protein Interaction Mapping, rabies virus, RHABDO, Two-Hybrid System Techniques, Viral Structural Proteins, VIRO, Virus Replication.
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Principales publications avant 2015

- 1) Ferlin A, Raux H, Baquero E, Lepault J, Gaudin Y. Characterization of pH sensitive molecular switches triggering the structural transition of VSV Glycoprotein from the post- toward the pre-fusion state. J Virol. 2014. Sep 10. [Epub ahead of print]

- 2) Baquero E, Albertini AA, Vachette P, Lepault J, Bressanelli S, Gaudin Y. Intermediate conformations during viral fusion glycoprotein structural transition. Curr Opin Virol. 2013. 3:143-50.

- 3) Albertini AA, Mérigoux C, Libersou S, Madiona K, Bressanelli S, Roche S, Lepault J, Melki R, Vachette P, Gaudin Y. Characterization of monomeric intermediates during VSV glycoprotein structural transition. PLoS Pathog. 2012. 8:e1002556.

- 4) Lahaye X, Vidy A, Fouquet B, Blondel D. Hsp70 protein positively regulates rabies virus infection. J Virol. 2012. 86:4743-51.

- 5) Fort P, Albertini A, Van-Hua A, Berthomieu A, Roche S, Delsuc F, Pasteur N, Capy P, Gaudin Y, Weill M. Fossil rhabdoviral sequences integrated into arthropod genomes : ontogeny, evolution, and potential functionality. Mol Biol Evol. 2012. 29:381-90.

- 6) Raux H, Obiang L, Richard N, Harper F, Blondel D, Gaudin Y. The matrix protein of vesicular stomatitis virus binds dynamin for efficient viral assembly. J Virol. 2010. 84:12609-18.

- 7) Libersou S, Albertini AA, Ouldali M, Maury V, Maheu C, Raux H, de Haas F, Roche S, Gaudin Y, Lepault J. Distinct structural rearrangements of the VSV glycoprotein drive membrane fusion. J Cell Biol. 2010. 191:199-210.

- 8) Blondel D, Kheddache S, Lahaye X, Dianoux L, Chelbi-Alix MK. Resistance to rabies virus infection conferred by the PMLIV isoform. J Virol. 2010. 84:10719-26.

- 9) Lahaye X, Vidy A, Pomier C, Obiang L, Harper F, Gaudin Y, Blondel D. Functional characterization of Negri bodies (NBs) in rabies virus-infected cells : Evidence that NBs are sites of viral transcription and replication. J Virol. 2009. 83:7948-58.

- 10) Roche S, Albertini AA, Lepault J, Bressanelli S, Gaudin Y. Structures of vesicular stomatitis virus glycoprotein : membrane fusion revisited. Cell Mol Life Sci. 2008. 65:1716-28. Review.

- 11) Vidy A, El Bougrini J, Chelbi-Alix MK, Blondel D. The nucleocytoplasmic rabies virus P protein counteracts interferon signaling by inhibiting both nuclear accumulation and DNA binding of STAT1. J Virol. 2007. 81:4255-63.

- 12) Weissenhorn W, Hinz A, Gaudin Y. Virus membrane fusion. FEBS Lett. 2007. 581:2150-5. Review.

- 13) Roche S, Rey FA, Gaudin Y, Bressanelli S. Structure of the prefusion form of the vesicular stomatitis virus glycoprotein G. Science. 2007. 315:843-8.

- 14) Roche S, Bressanelli S, Rey FA, Gaudin Y. Crystal structure of the low-pH form of the vesicular stomatitis virus glycoprotein G. Science. 2006. 313:187-91.

- 15) Vidy A, Chelbi-Alix M, Blondel D. Rabies virus P protein interacts with STAT1 and inhibits interferon signal transduction pathways. J Virol. 2005. 79:14411-20.

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