EWI-2wint promotes CD81 clustering that abrogates Hepatitis C Virus entry.
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 CD81 molecules 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.
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.
Julie
J
Potel
Patrice
P
Rassam
Claire
C
Montpellier
Laura
L
Kaestner
Elisabeth
E
Werkmeister
Birke A
BA
Tews
Cyril
C
Couturier
Costin-Ioan
CI
Popescu
Thomas F
TF
Baumert
Eric
E
Rubinstein
Jean
J
Dubuisson
Pierre-Emmanuel
PE
Milhiet
Laurence
L
Cocquerel
D015703:Antigens, CD / Q000378:metabolism*; D002460:Cell Line; D016174:Hepacivirus / Q000502:physiology*; D022781:Hepatocytes / Q000821:virology; D054884:Host-Pathogen Interactions; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D011991:Receptors, Virus / Q000378:metabolism*; D060225:Tetraspanin 28 / Q000378:metabolism*; D053586:Virus Internalization
D015703:Antigens, CD; C440028:IGSF8 protein, human; D008565:Membrane Proteins; D011991:Receptors, Virus; D060225:Tetraspanin 28