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Distinct roles in folding, CD81 receptor binding and viral entry for conserved histidine residues of hepatitis C virus glycoprotein E1 and E2.

Boo I, Tewierek K, Douam F, Lavillette D, Poumbourios P, Drummer HE
*Macfarlane Burnet Institute for Medical Research and Public Health, 85 Commerical Road, Melbourne, Victoria 3004, Australia.

Abstract

The protonation of histidine in acidic environments underpins its role in regulating the function of pH-sensitive proteins. For pH-sensitive viral fusion proteins, histidine protonation in the endosome leads to the activation of their membrane fusion function. The HCV (hepatitis C virus) glycoprotein E1-E2 heterodimer mediates membrane fusion within the endosome, but the roles of conserved histidine residues in the formation of a functional heterodimer and in sensing pH changes is unknown. We examined the functional roles of conserved histidine residues located within E1 and E2. The E1 mutations, H222A/R, H298R and H352A, disrupted E1-E2 heterodimerization and reduced virus entry. A total of five out of six histidine residues located within the E2 RBD (receptor-binding domain) were important for the E2 fold, and their substitution with arginine or alanine caused aberrant heterodimerization and/or CD81 binding. Distinct roles in E1-E2 heterodimerization and in virus entry were identified for His691 and His693 respectively within the membrane-proximal stem region. Viral entry and cell-cell fusion at neutral and low pH values were enhanced with H445R, indicating that the protonation state of His445 is a key regulator of HCV fusion. However, H445R did not overcome the block to virus entry induced by bafilomycin A1, indicating a requirement for an endosomal activation trigger in addition to acidic pH.

This work was supported by the National Health and Medical Research Council and the Agence Nationale de Recherche pour le SIDA et les hepatites virales’ (ANRS), the FINOVI foundation, Alliance funding. H.E.D. is supported by an RD Wright Fellowship. F.D. is a fellow of the French Ministry of Research.

Publisher’s full-text pdf of this article is available at:

http://www.biochemj.org/content/ppbiochemj/443/1/85.full.pdf

Publication

  • Journal: The Biochemical Journal
  • Published: 01/04/2012
  • Volume: 443
  • Issue: 1
  • Pagination: 85-94

Authors

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