1887

Abstract

The human immunodeficiency virus (HIV) gene encodes a 27 kDa myristoylated cytosolic protein that has an important role in the pathogenesis of AIDS. One function of Nef is the down-regulation of CD4 and MHC class I surface molecules in HIV-infected cells. Nef directly isolated from an infected individual (KS2), who could be defined as a long-term non-progressor, was compared with Nef from a standard laboratory strain, HIV-1 NL4-3. KS2 Nef protein was characterized by its lowered ability to down-regulate CD4, while still maintaining the ability to down-regulate MHC class I. The ability of KS2 Nef to down-regulate CD4 was more prominent when CD4 was measured 2–3 days after transfer of the gene to the target cells, and also when the effect was measured in CD4-enriched primary T cells. The amino acid sequence analysis indicated that the most notable feature of KS2 Nef was lack of the two glutamic acids: the EE region. When the EE region was added to KS2 Nef, the CD4 down-regulation ability was increased almost to the level of NL4-3 Nef. Conversely, when the EE region was deleted from NL4-3, its CD4 down-regulation ability was dramatically impaired. These data suggested that the EE region plays an important role(s) in the down-regulation of CD4 by Nef protein and also that primary sequences could be very useful in identifying the original biological functions of Nef .

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.79803-0
2004-06-01
2020-09-24
Loading full text...

Full text loading...

/deliver/fulltext/jgv/85/6/vir851451.html?itemId=/content/journal/jgv/10.1099/vir.0.79803-0&mimeType=html&fmt=ahah

References

  1. Arora V. K., Fredericksen B. L., Garcia J. V. 2002; Nef: agent of cell subversion. Microbes Infect 4:189–199 [CrossRef]
    [Google Scholar]
  2. Asamitsu K., Morishima T., Tsuchie H., Kurimura T., Okamoto T. 1999; Conservation of the central proline-rich (PxxP) motifs of human immunodeficiency virus type 1 Nef protein during the disease progression in two hemophiliac patients. FEBS Lett 459:399–404 [CrossRef]
    [Google Scholar]
  3. Benichou S., Bomsel M., Bodeus M., Durand H., Doute M., Letourneur F., Camonis J., Benarous R. 1994; Physical interaction of the HIV-1 Nef protein with beta-COP, a component of non-clathrin-coated vesicles essential for membrane traffic. J Biol Chem 269:30073–30076
    [Google Scholar]
  4. Benson R. E., Sanfridson A., Ottinger J. S., Doyle C., Cullen B. R. 1993; Downregulation of cell-surface CD4 expression by simian immunodeficiency virus Nef prevents viral super infection. J Exp Med 177:1561–1566 [CrossRef]
    [Google Scholar]
  5. Bresnahan P. A., Yonemoto W., Ferrell S., Williams-Herman D., Geleziunas R., Greene W. C. 1998; A dileucine motif in HIV-1 Nef acts as an internalization signal for CD4 downregulation and binds the AP-1 clathrin adaptor. Curr Biol 8:1235–1238 [CrossRef]
    [Google Scholar]
  6. Collins K. L., Chen B. K., Kalams S. A., Walker B. D., Baltimore D. 1998; HIV-1 Nef protein protects infected primary cells against killing by cytotoxic T lymphocytes. Nature 391:397–401 [CrossRef]
    [Google Scholar]
  7. Craig H. M., Pandori M. W., Guatelli J. C. 1998; Interaction of HIV-1 Nef with the cellular dileucine-based sorting pathway is required for CD4 down-regulation and optimal viral infectivity. Proc Natl Acad Sci U S A 95:11229–11234 [CrossRef]
    [Google Scholar]
  8. Garcia J. V., Miller A. D. 1991; Serine phosphorylation-independent downregulation of cell-surface CD4 by nef. Nature 350:508–511 [CrossRef]
    [Google Scholar]
  9. Geyer M., Peterlin B. M. 2001; Domain assembly, surface accessibility and sequence conservation in full length HIV-1 Nef. FEBS Lett 496:91–95 [CrossRef]
    [Google Scholar]
  10. Geyer M., Munte C. E., Schorr J., Kellner R., Kalbitzer H. R. 1999; Structure of the anchor-domain of myristoylated and non-myristoylated HIV-1 Nef protein. J Mol Biol 289:123–138 [CrossRef]
    [Google Scholar]
  11. Geyer M., Fackler O. T., Peterlin B. M. 2001; Structure–function relationships in HIV-1 Nef. EMBO Rep 2:580–585 [CrossRef]
    [Google Scholar]
  12. Greenberg M., DeTulleo L., Rapoport I., Skowronski J., Kirchhausen T. 1998a; A dileucine motif in HIV-1 Nef is essential for sorting into clathrin-coated pits and for downregulation of CD4. Curr Biol 8:1239–1242 [CrossRef]
    [Google Scholar]
  13. Greenberg M. E., Iafrate A. J., Skowronski J. 1998b; The SH3 domain-binding surface and an acidic motif in HIV-1 Nef regulate trafficking of class I MHC complexes. EMBO J 17:2777–2789 [CrossRef]
    [Google Scholar]
  14. Grzesiek S., Stahl S. J., Wingfield P. T., Bax A. 1996; The CD4 determinant for downregulation by HIV-1 Nef directly binds to Nef. Mapping of the Nef binding surface by NMR. Biochemistry 35:10256–10261 [CrossRef]
    [Google Scholar]
  15. Guy B., Kieny M. P., Riviere Y., Le Peuch C., Dott K., Girard M., Montagnier L., Lecocq J. P. 1987; HIV F/3′ orf encodes a phosphorylated GTP-binding protein resembling an oncogene product. Nature 330:266–269 [CrossRef]
    [Google Scholar]
  16. Janvier K., Craig H., Le Gall S., Benarous R., Guatelli J., Schwartz O., Benichou S. 2001; Nef-induced CD4 downregulation: a diacidic sequence in human immunodeficiency virus type 1 Nef does not function as a protein sorting motif through direct binding to beta-COP. J Virol 75:3971–3976 [CrossRef]
    [Google Scholar]
  17. Kestler H. W. III, Ringler D. J., Mori K., Panicali D. L., Sehgal P. K., Daniel M. D., Desrosiers R. C. 1991; Importance of the nef gene for maintenance of high virus loads and for development of AIDS. Cell 65:651–662 [CrossRef]
    [Google Scholar]
  18. Kirchhoff F., Greenough T. C., Brettler D. B., Sullivan J. L., Desrosiers R. C. 1995; Brief report: absence of intact nef sequences in a long-term survivor with nonprogressive HIV-1 infection. N Engl J Med 332:228–232 [CrossRef]
    [Google Scholar]
  19. Lama J., Mangasarian A., Trono D. 1999; Cell-surface expression of CD4 reduces HIV-1 infectivity by blocking Environ incorporation in a Nef- and Vpu-inhibitable manner. Curr Biol 9:622–631 [CrossRef]
    [Google Scholar]
  20. Lee J. T., Yu S. S., Han E., Kim S. 2004; Engineering the splice acceptor for improved gene expression and viral titer in an MLV-based retroviral vector. Gene Ther 11:94–99 [CrossRef]
    [Google Scholar]
  21. Mangasarian A., Piguet V., Wang J. K., Chen Y. L., Trono D. 1999; Nef-induced CD4 and major histocompatibility complex class I (MHC-I) down-regulation are governed by distinct determinants: N-terminal alpha helix and proline repeat of Nef selectively regulate MHC-I trafficking. J Virol 73:1964–1973
    [Google Scholar]
  22. Mariani R., Skowronski J. 1993; CD4 down-regulation by nef alleles isolated from human immunodeficiency virus type 1-infected individuals. Proc Natl Acad Sci U S A 90:5549–5553 [CrossRef]
    [Google Scholar]
  23. Nunn M. F., Marsh J. W. 1996; Human immunodeficiency virus type 1 Nef associates with a member of the p21-activated kinase family. J Virol 70:6157–6161
    [Google Scholar]
  24. Parnes J. R., Seong R. H. 1994; CD4 and CD8 in T cell lineage commitment: alterations induced by expression of a CD8/CD4 chimeric transgene. Semin Immunol 6:221–229 [CrossRef]
    [Google Scholar]
  25. Piguet V., Gu F., Foti M., Demaurex N., Gruenberg J., Carpentier J. L., Trono D. 1999; Nef-induced CD4 degradation: a diacidic-based motif in Nef functions as a lysosomal targeting signal through the binding of beta-COP in endosomes. Cell 97:63–73 [CrossRef]
    [Google Scholar]
  26. Piguet V., Wan L., Borel C., Mangasarian A., Demaurex N., Thomas G., Trono D. 2000; HIV-1 Nef protein binds to the cellular protein PACS-1 to downregulate class I major histocompatibility complexes. Nat Cell Biol 2:163–167 [CrossRef]
    [Google Scholar]
  27. Ross T. M., Oran A. E., Cullen B. R. 1999; Inhibition of HIV-1 progeny virion release by cell-surface CD4 is relieved by expression of the viral Nef protein. Curr Biol 9:613–621 [CrossRef]
    [Google Scholar]
  28. Schwartz O., Marechal V., Le Gall S., Lemonnier F., Heard J. M. 1996; Endocytosis of major histocompatibility complex class I molecules is induced by the HIV-1 Nef protein. Nat Med 2:338–342 [CrossRef]
    [Google Scholar]
  29. Soneoka Y., Cannon P. M., Ramsdale E. E., Griffiths J. C., Romano G., Kingsman S. M., Kingsman A. J. 1995; A transient three-plasmid expression system for the production of high titer retroviral vectors. Nucleic Acids Res 23:628–633 [CrossRef]
    [Google Scholar]
  30. Stoddart C. A., Geleziunas R., Ferrell S. 8 other authors 2003; Human immunodeficiency virus type 1 Nef-mediated downregulation of CD4 correlates with Nef enhancement of viral pathogenesis. J Virol 77:2124–2133 [CrossRef]
    [Google Scholar]
  31. Wysocki L. J., Sato V. L. 1978; “Panning” for lymphocytes: a method for cell selection. Proc Natl Acad Sci U S A 75:2844–2848 [CrossRef]
    [Google Scholar]
  32. Yoon K., Jeong J. G., Yoon H., Lee J. S., Kim S. 2001; Differential effects of primary human immunodeficiency virus type 1 nef sequences on downregulation of CD4 and MHC class I. Biochem Biophys Res Commun 284:638–642 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.79803-0
Loading
/content/journal/jgv/10.1099/vir.0.79803-0
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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