EWI-2wint promotes CD81 clustering that abrogates Hepatitis C Virus entry

Authors

  • Julie Potel,

    1. Hepatitis C Laboratory, Center for Infection and Immunity of Lille, University Lille Nord de France, CNRS-UMR8204, Inserm-U1019, Pasteur Institute of Lille, Lille, France
    Search for more papers by this author
  • Patrice Rassam,

    1. Centre de Biochimie Structurale, CNRS-UMR5048, Inserm-U1054, Université Sud de France, Montpellier, France
    Search for more papers by this author
  • Claire Montpellier,

    1. Hepatitis C Laboratory, Center for Infection and Immunity of Lille, University Lille Nord de France, CNRS-UMR8204, Inserm-U1019, Pasteur Institute of Lille, Lille, France
    Search for more papers by this author
  • Laura Kaestner,

    1. Hepatitis C Laboratory, Center for Infection and Immunity of Lille, University Lille Nord de France, CNRS-UMR8204, Inserm-U1019, Pasteur Institute of Lille, Lille, France
    Search for more papers by this author
  • Elisabeth Werkmeister,

    1. Bio Imaging Center Lille-Nord de France, Lille, France
    Search for more papers by this author
  • Birke A. Tews,

    1. Hepatitis C Laboratory, Center for Infection and Immunity of Lille, University Lille Nord de France, CNRS-UMR8204, Inserm-U1019, Pasteur Institute of Lille, Lille, France
    Current affiliation:
    1. Institut für Immunologie Friedrich-Loeffler Institut, Greifswald-Insel Riems, Germany
    Search for more papers by this author
  • Cyril Couturier,

    1. Inserm-U761, Université Lille 2, Lille, France
    Search for more papers by this author
  • Costin-Ioan Popescu,

    1. Institute of Biochemistry of the Romanian Academy, Bucharest, Romania
    Search for more papers by this author
  • Thomas F. Baumert,

    1. Inserm-U748, Université de Strasbourg, Pôle Hépato-digestif-Hôpitaux Universitaires de Strasbourg, Strasbourg, France
    Search for more papers by this author
  • Eric Rubinstein,

    1. Inserm-U1004, Institut André Lwoff, Université Paris-Sud, Villejuif, France
    Search for more papers by this author
  • Jean Dubuisson,

    1. Hepatitis C Laboratory, Center for Infection and Immunity of Lille, University Lille Nord de France, CNRS-UMR8204, Inserm-U1019, Pasteur Institute of Lille, Lille, France
    Search for more papers by this author
  • Pierre-Emmanuel Milhiet,

    1. Centre de Biochimie Structurale, CNRS-UMR5048, Inserm-U1054, Université Sud de France, Montpellier, France
    Search for more papers by this author
  • Laurence Cocquerel

    Corresponding author
    1. Hepatitis C Laboratory, Center for Infection and Immunity of Lille, University Lille Nord de France, CNRS-UMR8204, Inserm-U1019, Pasteur Institute of Lille, Lille, France
    • Institut für Immunologie Friedrich-Loeffler Institut, Greifswald-Insel Riems, Germany
    Search for more papers by this author

  • During the revision of the manuscript, another study showing that HCV exploits the dynamic properties of CD81 for its entry has been published (Harris et al., 2012), which is in accordance with our conclusions.

For correspondence. E-mail laurence.cocquerel@ibl.fr; Tel. (+33) 3 20 87 11 62; Fax (+33) 3 20 87 12 01.

Summary

CD81 is a major receptor for Hepatitis C Virus (HCV). It belongs to the tetraspanin family whose members form dynamic clusters with numerous partner proteins and with one another, forming tetraspanin-enriched areas in the plasma membrane. In our study, we combined single-molecule microscopy and biochemistry experiments to investigate the clustering and membrane behaviour of CD81 in the context of cells expressing EWI-2wint, a natural inhibitor of HCV entry. Interestingly, we found that EWI-2wint reduces the global diffusion of CD81 molecules due to a decrease of the diffusion rate of mobile CD81molecules and an increase in the proportion of confined molecules. Indeed, we demonstrated that EWI-2wint promotes CD81 clustering and confinement in CD81-enriched areas. In addition, we showed that EWI-2wint influences the colocalization of CD81 with Claudin-1 – a co-receptor required for HCV entry. Together, our results indicate that a change in membrane partitioning of CD81 occurs in the presence of EWI-2wint. This study gives new insights on the mechanism by which HCV enters into its target cells, namely by exploiting the dynamic properties of CD81.

Ancillary