Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1991 Oct;10(10):2747–2755. doi: 10.1002/j.1460-2075.1991.tb07823.x

Theoretical and functional analysis of the SIV fusion peptide.

M Horth 1, B Lambrecht 1, M C Khim 1, F Bex 1, C Thiriart 1, J M Ruysschaert 1, A Burny 1, R Brasseur 1
PMCID: PMC452983  PMID: 1915259

Abstract

The fusion domain of simian immunodeficiency virus (SIV) envelope glycoproteins is a hydrophobic region located at the amino-terminal extremity of the transmembrane protein (gp32). Assuming an alpha helical structure for the SIV fusogenic domain of gp32 in a lipid environment, theoretical studies have predicted that the fusion peptide would insert obliquely in the lipid bilayer. This oblique insertion could be an initial step of the fusion process by disorganizing locally the structure of the lipid bilayer. We have tested this hypothesis by selectively mutagenizing the SIV gp160 expressed via a vaccinia virus vector, to alter the theoretical angle of insertion of the fusion peptide. The fusogenic activity of the wild-type and mutant glycoproteins was tested after infection of T4 lymphocytic cell lines by the recombinant vaccinia virus, and measure of syncytia formation. Mutations that modified the oblique orientation reduced the fusogenic activity. In contrast, mutations that conserve the oblique orientation did not alter the fusogenic properties. Our results support the hypothesis that oblique orientation is important for fusogenic activity.

Full text

PDF
2747

Images in this article

2749Image
on p.2749
2750Image
on p.2750

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bosch M. L., Earl P. L., Fargnoli K., Picciafuoco S., Giombini F., Wong-Staal F., Franchini G. Identification of the fusion peptide of primate immunodeficiency viruses. Science. 1989 May 12;244(4905):694–697. doi: 10.1126/science.2541505. [DOI] [PubMed] [Google Scholar]
  2. Brasseur R., De Loof H., Ruysschaert J. M., Rosseneu M. Conformational analysis of lipid-associating proteins in a lipid environment. Biochim Biophys Acta. 1988 Aug 4;943(1):95–102. doi: 10.1016/0005-2736(88)90350-1. [DOI] [PubMed] [Google Scholar]
  3. Brasseur R., Lorge P., Goormaghtigh E., Ruysschaert J. M., Espion D., Burny A. The mode of insertion of the paramyxovirus F1 N-terminus into lipid matrix, an initial step in host cell/virus fusion. Virus Genes. 1988 Jul;1(4):325–332. doi: 10.1007/BF00257096. [DOI] [PubMed] [Google Scholar]
  4. Brasseur R., Ruysschaert J. M. Conformation and mode of organization of amphiphilic membrane components: a conformational analysis. Biochem J. 1986 Aug 15;238(1):1–11. doi: 10.1042/bj2380001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brasseur R., Vandenbranden M., Cornet B., Burny A., Ruysschaert J. M. Orientation into the lipid bilayer of an asymmetric amphipathic helical peptide located at the N-terminus of viral fusion proteins. Biochim Biophys Acta. 1990 Nov 16;1029(2):267–273. doi: 10.1016/0005-2736(90)90163-i. [DOI] [PubMed] [Google Scholar]
  6. Chakrabarti S., Brechling K., Moss B. Vaccinia virus expression vector: coexpression of beta-galactosidase provides visual screening of recombinant virus plaques. Mol Cell Biol. 1985 Dec;5(12):3403–3409. doi: 10.1128/mcb.5.12.3403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dalgleish A. G., Beverley P. C., Clapham P. R., Crawford D. H., Greaves M. F., Weiss R. A. The CD4 (T4) antigen is an essential component of the receptor for the AIDS retrovirus. Nature. 1984 Dec 20;312(5996):763–767. doi: 10.1038/312763a0. [DOI] [PubMed] [Google Scholar]
  8. Eisenberg D., Weiss R. M., Terwilliger T. C. The helical hydrophobic moment: a measure of the amphiphilicity of a helix. Nature. 1982 Sep 23;299(5881):371–374. doi: 10.1038/299371a0. [DOI] [PubMed] [Google Scholar]
  9. Felser J. M., Klimkait T., Silver J. A syncytia assay for human immunodeficiency virus type I (HIV-I) envelope protein and its use in studying HIV-I mutations. Virology. 1989 Jun;170(2):566–570. doi: 10.1016/0042-6822(89)90448-0. [DOI] [PubMed] [Google Scholar]
  10. Freed E. O., Myers D. J., Risser R. Characterization of the fusion domain of the human immunodeficiency virus type 1 envelope glycoprotein gp41. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4650–4654. doi: 10.1073/pnas.87.12.4650. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gallaher W. R. Detection of a fusion peptide sequence in the transmembrane protein of human immunodeficiency virus. Cell. 1987 Jul 31;50(3):327–328. doi: 10.1016/0092-8674(87)90485-5. [DOI] [PubMed] [Google Scholar]
  12. Goormaghtigh E., Martin I., Vandenbranden M., Brasseur R., Ruysschaert J. M. Secondary structure and orientation of a chemically synthesized mitochondrial signal sequence in phospholipid bilayers. Biochem Biophys Res Commun. 1989 Jan 31;158(2):610–616. doi: 10.1016/s0006-291x(89)80093-2. [DOI] [PubMed] [Google Scholar]
  13. Harter C., James P., Bächi T., Semenza G., Brunner J. Hydrophobic binding of the ectodomain of influenza hemagglutinin to membranes occurs through the "fusion peptide". J Biol Chem. 1989 Apr 15;264(11):6459–6464. [PubMed] [Google Scholar]
  14. Klatzmann D., Champagne E., Chamaret S., Gruest J., Guetard D., Hercend T., Gluckman J. C., Montagnier L. T-lymphocyte T4 molecule behaves as the receptor for human retrovirus LAV. Nature. 1984 Dec 20;312(5996):767–768. doi: 10.1038/312767a0. [DOI] [PubMed] [Google Scholar]
  15. Kornfeld H., Riedel N., Viglianti G. A., Hirsch V., Mullins J. I. Cloning of HTLV-4 and its relation to simian and human immunodeficiency viruses. Nature. 1987 Apr 9;326(6113):610–613. doi: 10.1038/326610a0. [DOI] [PubMed] [Google Scholar]
  16. Kowalski M., Potz J., Basiripour L., Dorfman T., Goh W. C., Terwilliger E., Dayton A., Rosen C., Haseltine W., Sodroski J. Functional regions of the envelope glycoprotein of human immunodeficiency virus type 1. Science. 1987 Sep 11;237(4820):1351–1355. doi: 10.1126/science.3629244. [DOI] [PubMed] [Google Scholar]
  17. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  18. Lasky L. A., Nakamura G., Smith D. H., Fennie C., Shimasaki C., Patzer E., Berman P., Gregory T., Capon D. J. Delineation of a region of the human immunodeficiency virus type 1 gp120 glycoprotein critical for interaction with the CD4 receptor. Cell. 1987 Sep 11;50(6):975–985. doi: 10.1016/0092-8674(87)90524-1. [DOI] [PubMed] [Google Scholar]
  19. Mackett M., Smith G. L., Moss B. General method for production and selection of infectious vaccinia virus recombinants expressing foreign genes. J Virol. 1984 Mar;49(3):857–864. doi: 10.1128/jvi.49.3.857-864.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Marsh M., Helenius A. Virus entry into animal cells. Adv Virus Res. 1989;36:107–151. doi: 10.1016/S0065-3527(08)60583-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Martin I., Defrise-Quertain F., Mandieau V., Nielsen N. M., Saermark T., Burny A., Brasseur R., Ruysschaert J. M., Vandenbranden M. Fusogenic activity of SIV (simian immunodeficiency virus) peptides located in the GP32 NH2 terminal domain. Biochem Biophys Res Commun. 1991 Mar 29;175(3):872–879. doi: 10.1016/0006-291x(91)91646-t. [DOI] [PubMed] [Google Scholar]
  22. McCune J. M., Rabin L. B., Feinberg M. B., Lieberman M., Kosek J. C., Reyes G. R., Weissman I. L. Endoproteolytic cleavage of gp160 is required for the activation of human immunodeficiency virus. Cell. 1988 Apr 8;53(1):55–67. doi: 10.1016/0092-8674(88)90487-4. [DOI] [PubMed] [Google Scholar]
  23. Rafalski M., Lear J. D., DeGrado W. F. Phospholipid interactions of synthetic peptides representing the N-terminus of HIV gp41. Biochemistry. 1990 Aug 28;29(34):7917–7922. doi: 10.1021/bi00486a020. [DOI] [PubMed] [Google Scholar]
  24. Taylor J. W., Ott J., Eckstein F. The rapid generation of oligonucleotide-directed mutations at high frequency using phosphorothioate-modified DNA. Nucleic Acids Res. 1985 Dec 20;13(24):8765–8785. doi: 10.1093/nar/13.24.8765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Verkleij A. J. Lipidic intramembranous particles. Biochim Biophys Acta. 1984 Jan 27;779(1):43–63. doi: 10.1016/0304-4157(84)90003-0. [DOI] [PubMed] [Google Scholar]
  26. Wharton S. A., Martin S. R., Ruigrok R. W., Skehel J. J., Wiley D. C. Membrane fusion by peptide analogues of influenza virus haemagglutinin. J Gen Virol. 1988 Aug;69(Pt 8):1847–1857. doi: 10.1099/0022-1317-69-8-1847. [DOI] [PubMed] [Google Scholar]
  27. White J. M. Viral and cellular membrane fusion proteins. Annu Rev Physiol. 1990;52:675–697. doi: 10.1146/annurev.ph.52.030190.003331. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES