1887

Abstract

Simian immunodeficiency virus (SIV) and human immunodeficiency virus type 2 (HIV-2) encode the accessory viral protein X (Vpx) known to be incorporated into virions in amounts comparable to those of the Gag proteins. The localization of Vpx within SIV-infected HUT-78 cells and SIV virions was studied by immunoelectron microscopy. Vpx appeared to be associated with extracellular virions as well as budding viral particles at the surface of infected cells. Immunolabelling of purified viral cores suggested that Vpx was a component of the amorphous material surrounding the core structure. Furthermore, a detergent insoluble fraction containing SIV core proteins was devoid of Vpx. To investigate the protein requirement for packaging of Vpx, BHK-21 cells were co-infected with vaccinia virus recombinants encoding Vpx and other SIV proteins able to assemble into virus-like particles. Analysis by immunoprecipitation of the extracellular particulate material as well as immunoelectron microscopy demonstrated that co-expression of Vpx with the Pr56 polyprotein was sufficient for the formation of pseudo-virions containing Vpx. Virus-like particles that appeared upon expression of p16 did not contain Vpx. The results suggest that Vpx is packaged into viral particles through its binding to the Gag polyprotein. The precise positioning of Vpx within the space separating the viral envelope from the core structure is postulated to result from the reorganization of viral proteins that occurs upon Gag polyprotein cleavage and budding.

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1994-11-01
2022-05-21
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References

  1. Chakrabarti L., Guyader M., Alizon M., Daniel M. D., Desrosiers R. C., Tiollais P., Sonigo P. 1987; Sequence of simian immunodeficiency virus from macaque and its relationship to other human and simian retroviruses. Nature; London: 328543–547
    [Google Scholar]
  2. Chrystie I. L., Almeida J. D. 1989; Recovery of antigenically reactive HIV-2 cores. Journal of Medical Virology 27:188–195
    [Google Scholar]
  3. Cohen E., Dehni G., Sodroski J. G., Haseltine W. A. 1990; Human immunodeficiency virus Vpr product is a virion-associated regulatory protein. Journal of Virology 64:3097–3099
    [Google Scholar]
  4. Delchambre M., Gheysen D., Thines D., Thiriart C., Jacobs E., Verdin E., Horth M., Burny A., Bex F. 1989; The GAG precursor of simian immunodeficiency virus assembles into virus-like particles. EMBO Journal 8:2653–2660
    [Google Scholar]
  5. Desrosiers R. C. 1992; HIV with multiple gene deletions as a live attenuated vaccine for AIDS. AIDS Research and Human Retroviruses 8:387–393
    [Google Scholar]
  6. Franchini G., Gurgo C., Guo H. G., Gallo R. C., Collalti E., Fargnoli K. A., Hall L. F., Wong-Staal F., Reitz M. S.Jr 1987; Sequence of simian immunodeficiency virus and its re-lationship to the human immunodeficiency viruses. Nature; London: 328539–542
    [Google Scholar]
  7. Franchini G., Rusche J. R., O’keeffe T. J., Wong-Staal F. 1988; The human immunodeficiency virus type 2 (HIV-2) contains a novel gene encoding a 16 kD protein associated with mature virions. AIDS Research and Human Retroviruses 4:243–250
    [Google Scholar]
  8. Gheysen D., Jacobs E., Foresta F., Thiriart C., Francotte M., Thines D., De Wilde M. 1989; Assembly and release of HIV-1 precursor Pr55gag virus-like particles from recombinant baculovirus- infected cells. Cell 59:103–112
    [Google Scholar]
  9. Gonzales S. A., Affrachino J. L., Gelderblom H. R., Burny A. 1993; Assembly of the matrix protein of simian immunodeficiency virus into virus-like particles. Virology 194:548–556
    [Google Scholar]
  10. Grief C., Hockley D. J., Fromholc C. E., Kitchin P. A. 1989; The morphology of simian immunodeficiency virus as shown by negative staining electron microscopy. Journal of General Virology 70:2215–2219
    [Google Scholar]
  11. Grigoriev V. B., Kodochicov J. P., Rudneva I. A., Kamarov S. M., Escaig F., Fournier J. -G. 1989; Detection by immuno- gold technique of HIV antigens in Lowicryl ultrathin sections of infected cells. Pathology Research and Practice 184:494–497
    [Google Scholar]
  12. Guyader M., Emerman M., Sonigo P., Clavel F., Montagnier L., Alizon M. 1987; Genome organization and transactivation of the human immunodeficiency virus type 2. Nature; London: 326662–669
    [Google Scholar]
  13. Guyader M., Emerman M., Montagnier L., Peden K. 1989; VPX mutants of HIV-2 are infectious in established cell lines but display a severe defect in peripheral blood lymphocytes. EMBO Journal 8:1169–1175
    [Google Scholar]
  14. Haffar O., Garrigues J., Travis B., Moran P., Zarling J., Hu S. L. 1990; Human immunodeficiency virus-like, nonreplicating gag-env particles assemble in a recombinant vaccinia virus expression system. Journal of Virology 64:2653–2659
    [Google Scholar]
  15. Henderson L. E., Benveniste R. E., Sowder R., Copeland T.D, Oroszlan S. 1988a; Molecular characterization of Gag proteins from simian immunodeficiency virus (SIVMne). Journal of Virology 62:2587–2595
    [Google Scholar]
  16. Henderson L. E., Sowder R. C., Benveniste R. E., Oroszlan S. 1988b; Isolation and characterization of a novel protein (X-ORF product) from SIV and HIV-2. Science 241:199–201
    [Google Scholar]
  17. Hirsch V. M., Johnson P. R. 1994; Pathogenic diversity of simian immunodeficiency viruses. Virus Research 32:183–203
    [Google Scholar]
  18. Hu W., Vander Heyden N., Ratner L. 1989; Analysis of the function of viral protein X (Vpx) of HIV-2. Virology 173:624–630
    [Google Scholar]
  19. Kappes J. C., Conway J. A., Lee S. W., Shaw G. M., Hahn B. H. 1991; Human immunodeficiency virus type 2 VPX protein augments viral infectivity. Virology 184:197–209
    [Google Scholar]
  20. Kappes J. C., Parkin J. S., Conway J. A., Kim J., Brouillette C. G., Shaw G. M., Hahn B. H. 1993; Intracellular transport and virion incorporation of VPX requires interaction with other virus type-specific components. Virology 193:222–233
    [Google Scholar]
  21. Karacostas V., Nagashima K., Gonda M. A., Moss B. 1989; Human immunodeficiency virus-like particles produced by a vaccinia virus expression vector. Proceedings of the National Academy of Sciences U.S.A: 868964–8967
    [Google Scholar]
  22. Kieny M. P., Lathe R., Drillien R., Spehner D., Skory S., Schmitt D., Wiktor T., Koprowski H., Lecocq J. P. 1984; Expression of rabies virus glycoprotein from a recombinant vaccinia virus. Nature; London: 312163–166
    [Google Scholar]
  23. Lu Y. -L., Spearman P., Ratner L. 1994; Human immunodeficiency virus type 1 viral protein R localization in infected cells and virions. Journal of Virology 67:6542–6550
    [Google Scholar]
  24. Maniatis T., Fritsch E. F., Sambrook J. 1989 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  25. Marx P. A., Munn R. J., Joy K. I. 1988; Computer emulation of thin section electron microscopy predicts an envelope-associated icosahedral capsid for human immunodeficiency virus. Laboratory Investigation 58:112–118
    [Google Scholar]
  26. Paxton W., Connor R. I., Landau N. R. 1994; Incorporation of Vpr into human immunodeficiency virus type 1 virions: requirement for the p6 region of gag and mutational analysis. Journal of Virology 67:7229–7237
    [Google Scholar]
  27. Shibata R., Adachi A., Sakai H., Ishimoto A., Miura T., Hayami M. 1990; Mutational analysis of simian immunodeficiency virus from African green monkeys and human immunodeficiency virus type 2. Journal of Medical Primalology 19:217–225
    [Google Scholar]
  28. Shioda T., Shibuta H. 1990; Production of human immunodeficiency virus (HlV-l)-like particles from cells infected with recombinant vaccinia viruses carrying the gag gene of HIV. Virology 175:139–148
    [Google Scholar]
  29. South T. L., Blake P. R., Sowder R. C., Arthur L. O., Henderson L. E., Summers M. F. 1990; The nucleocapsid protein isolated from HIV-1 particles binds zinc and forms retroviral- type zinc fingers. Biochemistry 29:7786–7789
    [Google Scholar]
  30. Spehner D., Kirn A., Drillien R. 1991; Assembly of nucleocapsidlike structures in animal cells infected with a vaccinia virus recombinant encoding the measles virus nucleoprotein. Journal of Virology 65:6296–6300
    [Google Scholar]
  31. Tristem M., Marshall C., Karpas A., Petrik J., Hill F. 1990; Origin of VPX in lentiviruses. Nature; London: 347341–342
    [Google Scholar]
  32. Tristem M., Marshall C., Karpas A., Hill F. 1992; Evolution of the primate lentiviruses: evidence from VPX and VPR. EMBO Journal 11:3405–3412
    [Google Scholar]
  33. Vzorov A. N., Bukrinsky M. I., Grigoriev V. B., Tentsov Y. Y., Bukrinskaya A. G. 1991; Highly immunogenic human immunodeficiency virus-like particles are produced by recombinant vaccinia virus-infected cells. AIDS Research and Human Retroviruses 7:29–36
    [Google Scholar]
  34. Whitehouse R. S., Benichou J. C., Couture-Tosi E., Schenkman S., Ryter A. 1984; Immunolabelling of lambda receptor protein (lam B) on thin sections of E. coli embedded in Lowicryl. Biology of the Cell 51:389–394
    [Google Scholar]
  35. Yu X. F., Ito S., Essex M., Lee T. H. 1988; A naturally immunogenic virion-associated protein specific for HIV-2 and SIV. Nature; London: 335262–265
    [Google Scholar]
  36. Yu X. F., Matsuda M., Essex M., Lee T. H. 1990; Open reading frame vpr of simian immunodeficiency virus encodes a virion- associated protein. Journal of Virology 64:5688–5693
    [Google Scholar]
  37. Yu X. F., Yu Q. C., Essex M., Lee T. H. 1991; The vpx gene of simian immunodeficiency virus facilitates efficient viral replication in fresh lymphocytes and macrophages. Journal of Virology 65:5088–5091
    [Google Scholar]
  38. Yu X. F., Matsuda Z., Yu Q. C., Lee T. H., Essex M. 1993; Vpx of simian immunodeficiency virus is localized primarily outside the virus core in mature virions. Journal of Virology 67:4386–4390
    [Google Scholar]
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