1887

Abstract

The E1E2 glycoprotein heterodimer of mediates viral entry. E2 attaches the virus to cellular receptors; however, the function of E1 is unknown. We tested the hypothesis that E1 is a truncated class II fusion protein. We mutated amino acids within a predicted fusion peptide (residues 276–286) and a truncated C-terminal stem-like motif, containing a membrane-proximal heptad-repeat sequence (residues 330–347). The fusion peptide mutation F285A abolished viral entry, while mutation of other hydrophobic residues had no effect. Alanine replacement of heptad-repeat residues blocked entry in three of five cases, whereas substitution with the helix breaker, Pro, led to loss of entry function in all cases. The mutations did not affect glycoprotein expression, heterodimerization with E2 or global folding, in contrast to the effects of mutations in the fusion motifs of prototypical class II fusion proteins. Our data suggest that E1 is unlikely to function in an analogous manner to other class II fusion glycoproteins.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.82567-0
2007-04-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/88/4/1144.html?itemId=/content/journal/jgv/10.1099/vir.0.82567-0&mimeType=html&fmt=ahah

References

  1. Allison S. L., Stiasny K., Stadler K., Mandl C. W., Heinz F. X. 1999; Mapping of functional elements in the stem-anchor region of tick-borne encephalitis virus envelope protein E. J Virol 73:5605–5612
    [Google Scholar]
  2. Allison S. L., Schalich J., Stiasny K., Mandl C. W., Heinz F. X. 2001; Mutational evidence for an internal fusion peptide in flavivirus envelope protein E. J Virol 75:4268–4275 [CrossRef]
    [Google Scholar]
  3. Bartosch B., Cosset F. L. 2006; Cell entry of hepatitis C virus. Virology 348:1–12 [CrossRef]
    [Google Scholar]
  4. Bartosch B., Dubuisson J., Cosset F. L. 2003; Infectious hepatitis C virus pseudo-particles containing functional E1–E2 envelope protein complexes. J Exp Med 197:633–642 [CrossRef]
    [Google Scholar]
  5. Blanchard E., Belouzard S., Goueslain L., Wakita T., Dubuisson J., Wychowski C., Rouille Y. 2006; Hepatitis C virus entry depends on clathrin-mediated endocytosis. J Virol 80:6964–6972 [CrossRef]
    [Google Scholar]
  6. Bressanelli S., Stiasny K., Allison S. L., Stura E. A., Duquerroy S., Lescar J., Heinz F. X., Rey F. A. 2004; Structure of a flavivirus envelope glycoprotein in its low-pH-induced membrane fusion conformation. EMBO J 23:728–738 [CrossRef]
    [Google Scholar]
  7. Chang D. K., Cheng S. F., Trivedi V. D., Lin K. L. 1999; Proline affects oligomerization of a coiled coil by inducing a kink in a long helix. J Struct Biol 128:270–279 [CrossRef]
    [Google Scholar]
  8. Codran A., Royer C., Jaeck D., Bastien-Valle M., Baumert T. F., Kieny M. P., Pereira C. A., Martin J. P. 2006; Entry of hepatitis C virus pseudotypes into primary human hepatocytes by clathrin-dependent endocytosis. J Gen Virol 87:2583–2593 [CrossRef]
    [Google Scholar]
  9. Delos S. E., Gilbert J. M., White J. M. 2000; The central proline of an internal viral fusion peptide serves two important roles. J Virol 74:1686–1693 [CrossRef]
    [Google Scholar]
  10. Drummer H. E., Poumbourios P. 2004; Hepatitis C virus glycoprotein E2 contains a membrane-proximal heptad repeat sequence that is essential for E1E2 glycoprotein heterodimerization and viral entry. J Biol Chem 279:30066–30072 [CrossRef]
    [Google Scholar]
  11. Drummer H. E., Wilson K. A., Poumbourios P. 2002; Identification of the hepatitis C virus e2 glycoprotein binding site on the large extracellular loop of CD81. J Virol 76:11143–11147 [CrossRef]
    [Google Scholar]
  12. Drummer H. E., Maerz A., Poumbourios P. 2003; Cell surface expression of functional hepatitis C virus E1 and E2 glycoproteins. FEBS Lett 546:385–390 [CrossRef]
    [Google Scholar]
  13. Drummer H. E., Boo I., Maerz A. L., Poumbourios P. 2006; A conserved Gly436-Trp-Leu-Ala-Gly-Leu-Phe-Tyr motif in hepatitis C virus glycoprotein e2 is a determinant of CD81 binding and viral entry. J Virol 80:7844–7853 [CrossRef]
    [Google Scholar]
  14. Garry R. F., Dash S. 2003; Proteomics computational analyses suggest that hepatitis C virus E1 and pestivirus E2 envelope glycoproteins are truncated class II fusion proteins. Virology 307:255–265 [CrossRef]
    [Google Scholar]
  15. He J., Landau N. R. 1995; Use of a novel human immunodeficiency virus type 1 reporter virus expressing human placental alkaline phosphatase to detect an alternative viral receptor. J Virol 69:4587–4592
    [Google Scholar]
  16. Hsu M., Zhang J., Flint M., Logvinoff C., Cheng-Mayer C., Rice C. M., McKeating J. A. 2003; Hepatitis C virus glycoproteins mediate pH-dependent cell entry of pseudotyped retroviral particles. Proc Natl Acad Sci U S A 100:7271–7276 [CrossRef]
    [Google Scholar]
  17. Keck Z. Y., Op De Beeck A., Hadlock K. G., Xia J., Li T. K., Dubuisson J., Foung S. K. 2004; Hepatitis C virus e2 has three immunogenic domains containing conformational epitopes with distinct properties and biological functions. J Virol 78:9224–9232 [CrossRef]
    [Google Scholar]
  18. Keck Z. Y., Li T. K., Xia J., Bartosch B., Cosset F. L., Dubuisson J., Foung S. K. 2005; Analysis of a highly flexible conformational immunogenic domain a in hepatitis C virus E2. J Virol 79:13199–13208 [CrossRef]
    [Google Scholar]
  19. Kielian M. 2006; Class II virus membrane fusion proteins. Virology 344:38–47 [CrossRef]
    [Google Scholar]
  20. Kielian M., Rey F. A. 2006; Virus membrane-fusion proteins: more than one way to make a hairpin. Nat Rev Microbiol 4:67–76 [CrossRef]
    [Google Scholar]
  21. Lavillette D., Bartosch B., Nourrisson D., Verney G., Cosset F. L., Penin F., Pecheur E. I. 2006; Hepatitis C virus glycoproteins mediate low pH-dependent membrane fusion with liposomes. J Biol Chem 281:3909–3917 [CrossRef]
    [Google Scholar]
  22. Lindenbach B. D., Evans M. J., Syder A. J., Wolk B., Tellinghuisen T. L., Liu C. C., Maruyama T., Hynes R. O., Burton D. R. other authors 2005; Complete replication of hepatitis C virus in cell culture. Science 309:623–626 [CrossRef]
    [Google Scholar]
  23. Modis Y., Ogata S., Clements D., Harrison S. C. 2004; Structure of the dengue virus envelope protein after membrane fusion. Nature 427:313–319 [CrossRef]
    [Google Scholar]
  24. Owsianka A. M., Timms J. M., Tarr A. W., Brown R. J., Hickling T. P., Szwejk A., Bienkowska-Szewczyk K., Thomson B. J., Patel A. H., Ball J. K. 2006; Identification of conserved residues in the e2 envelope glycoprotein of the hepatitis C virus that are critical for CD81 binding. J Virol 80:8695–8704 [CrossRef]
    [Google Scholar]
  25. Pacheco B., Gomez-Gutierrez J., Yelamos B., Delgado C., Roncal F., Albar J. P., Peterson D., Gavilanes F. 2006; Membrane-perturbing properties of three peptides corresponding to the ectodomain of hepatitis C virus E2 envelope protein. Biochim Biophys Acta 1758755–763 [CrossRef]
    [Google Scholar]
  26. Perez-Berna A. J., Moreno M. R., Guillen J., Bernabeu A., Villalain J. 2006; The membrane-active regions of the hepatitis C virus E1 and E2 envelope glycoproteins. Biochemistry 45:3755–3768 [CrossRef]
    [Google Scholar]
  27. Rey F. A., Heinz F. X., Mandl C., Kunz C., Harrison S. C. 1995; The envelope glycoprotein from tick-borne encephalitis virus at 2 Å resolution. Nature 375:291–298 [CrossRef]
    [Google Scholar]
  28. Skehel J. J., Wiley D. C. 2000; Receptor binding and membrane fusion in virus entry: the influenza hemagglutinin. Annu Rev Biochem 69:531–569 [CrossRef]
    [Google Scholar]
  29. Stiasny K., Heinz F. X. 2006; Flavivirus membrane fusion. J Gen Virol 87:2755–2766 [CrossRef]
    [Google Scholar]
  30. Tscherne D. M., Jones C. T., Evans M. J., Lindenbach B. D., McKeating J. A., Rice C. M. 2006; Time- and temperature-dependent activation of hepatitis C virus for low-pH-triggered entry. J Virol 80:1734–1741 [CrossRef]
    [Google Scholar]
  31. Yagnik A. T., Lahm A., Meola A., Roccasecca R. M., Ercole B. B., Nicosia A., Tramontano A. 2000; A model for the hepatitis C virus envelope glycoprotein E2. Proteins 40:355–366 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.82567-0
Loading
/content/journal/jgv/10.1099/vir.0.82567-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error