In silico directed mutagenesis identifies the CD 81/claudin‐1 hepatitis C virus receptor interface

Davis, C. , Harris, H. J., Hu, K., Drummer, H. E., McKeating, J. A., Mullins, J. G.L. and Balfe, P. (2012) In silico directed mutagenesis identifies the CD 81/claudin‐1 hepatitis C virus receptor interface. Cellular Microbiology, 14(12), pp. 1892-1903. (doi: 10.1111/cmi.12008) (PMID:22897233) (PMCID:PMC3549482)

Abstract

Hepatitis C virus (HCV) entry is dependent on host cell molecules tetraspanin CD81, scavenger receptor BI and tight junction proteins claudin-1 and occludin. We previously reported a role for CD81/claudin-1 receptor complexes in HCV entry; however, the molecular mechanism(s) driving association between the receptors is unknown. We explored the molecular interface between CD81 and claudin-1 using a combination of bioinformatic sequence-based modelling, site-directed mutagenesis and Fluorescent Resonance Energy Transfer (FRET) imaging methodologies. Structural modelling predicts the first extracellular loop of claudin-1 to have a flexible beta conformation and identifies a motif between amino acids 62–66 that interacts with CD81 residues T149, E152 and T153. FRET studies confirm a role for these CD81 residues in claudin-1 association and HCV infection. Importantly, mutation of these CD81 residues has minimal impact on protein conformation or HCVglycoprotein binding, highlighting a new functional domain of CD81 that is essential for virus entry.

• Item Type: Articles
• Additional Information: This work was supported by the MRC (Grants G0400802 and G0801976), The Wellcome Trust (Grant ME027881) and by the European Union ‘HEPACUTE (FP7)’ programme.
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Davis, Dr Chris
• Authors: Davis, C., Harris, H. J., Hu, K., Drummer, H. E., McKeating, J. A., Mullins, J. G.L., and Balfe, P.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity
• Journal Name: Cellular Microbiology
• Publisher: Wiley
• ISSN: 1462-5814
• ISSN (Online): 1462-5822
• Published Online: 25 September 2012