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A C-terminal domain of HIV-1 accessory protein Vpr is involved in penetration, mitochondrial dysfunction and apoptosis of human CD4+ lymphocytes

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Abstract

We have previously shown that expression of HIV-1 vpr in yeast results in cell growth arrest and structural defects, and identified a C-terminal domain of Vpr as being responsible for these effects in yeast.1 In this report we show that recombinant Vpr and C-terminal peptides of Vpr containing the conserved sequence HFRIGCRHSRIG caused permeabilization of CD4+ T lymphocytes, a dramatic reduction of mitochondrial membrane potential and finally cell death. Vpr and Vpr peptides containing the conserved sequence rapidly penetrated cells, co-localized with the DNA, and caused increased granularity and formation of dense apoptotic bodies. The above results suggest that Vpr treated cells undergo apoptosis and this was confirmed by demonstration of DNA fragmentation by the highly sensitive TUNEL assay. Our results, together with the demonstration of extracellular Vpr in HIV infected individuals,2,3 suggest the possibility that extracellular Vpr could contribute to the apoptotic death and depletion of bystander cells in lymphoid tissues4,5 during HIV infection.

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References

  1. Macreadie IG, Castelli LA, Hewish DR, Kirkpatrick A, Ward AC, Azad AA. A domain of human immunodeficiency virus type 1 Vpr containing repeated H(S/F)RIG amino acid motifs causes cell growth arrest and structural defects. Proc Natl Acad Sci USA 1995; 92: 2770-2774.

    Google Scholar 

  2. Levy DN, Refaeli Y, MacGregor RR, Weiner DB. Serum Vpr regulates productive infection and latency of human immunodeficiency virus type 1. Proc Natl Acad Sci USA 1994; 91: 10873-10877.

    Google Scholar 

  3. Levy DN, Refaeli Y, Weiner DB. Extracellular Vpr protein increases cellular permissiveness to human immunodeficiency virus replication and reactivates virus from latency. J Virol 1995; 69: 1243-1252.

    Google Scholar 

  4. Su L, Kaneshima H, Bonyhadi M, Salimi S, Kraft D, Rabin L, McCune JM. HIV-1-induced thymocyte depletion is associated with indirect cytopathicity and infection of progenitor cells in vivo. Immunity 1995; 2: 25-36

    Google Scholar 

  5. Finkel TH, Tudor-Williams G, Banda NK, et al. Apoptosis occurs predominantly in bystander cells and not in productively infected cells of HIV-and SIV-infected lymph nodes. Nature Med 1995; 1: 129-134.

    Google Scholar 

  6. Cohen EA, Dehni G, Sodrowski JG, Haseltine WA. Human immunodeficiency virus vpr product is a virion-associated regulatory protein. J Virol 1990; 64, 3097-3099.

    Google Scholar 

  7. Yuan X, Matsuda Z, Matsuda M, Essex M, Lee TH. Human immunodeficiency virus vpr gene encodes a virion-associated protein. AIDS Res Human Retroviruses 1990; 6: 1265-1271.

    Google Scholar 

  8. Yu XF, Matsuda M, Essex M, Lee TH. Open reading frame vpr of simian immunodeficiency virus encodes a virion-associated protein. J Virol 1990; 64: 5688-5693.

    Google Scholar 

  9. Heinzinger NK, Bukrinsky MI, Haggerty SA, et al. The Vpr protein of human immunodeficiency virus type 1 influences nuclear localization of viral nucleic acids in nondividing host cells. Proc Natl Acad Sci USA 1994; 91: 7311-7315.

    Google Scholar 

  10. Hattori N, Michaels F, Fargnoli K, Marcon L, Gallo RC, Franchini G. The human immunodeficiency virus type 2 vpr gene is essential for productive infection of human macrophages. Proc Natl Acad Sci USA 1990; 87: 8080-8084.

    Google Scholar 

  11. Balotta C, Lusso P, Crowley R, Gallo RC, Franchini G. Antisense phosphothioate oligonucleotides targeted to the vpr gene inhibit human immunodeficiency virus type 1 replication in primary macrophages. J Virol 1993; 67: 4409-4414.

    Google Scholar 

  12. Matsuda Z, Yu X, Yu QC, Lee TH, Essex M. A virion-specific inhibitory molecule with therapeutic potential for human immunodeficiency virus type 1. Proc Natl Acad Sci USA 1993; 90: 3544-3548.

    Google Scholar 

  13. Connor RI, Chen BK, Choe S, Landau NR. Vpr is required for efficient replication of human immunodeficiency virus type 1 in mononuclear phagocytes. Virology 1995; 206: 935-944.

    Google Scholar 

  14. Lang SM, Weeger M, Stahl-Henning C, et al. Importance of vpr for infection of rhesus monkeys with simian immunodeficiency virus. J Virol 1993; 67: 902-912.

    Google Scholar 

  15. Levy DN, Fernandes LS, Williams WV, Weiner DB. Induction of cell differentiation by human immunodeficiency virus 1 vpr. Cell 1993; 72: 541-550.

    Google Scholar 

  16. Rogel ME, Wu LI, Emerman M. The human immunodeficiency virus type 1 vpr gene prevents cell proliferation during chronic infection. J Virol 1995; 69: 882-888.

    Google Scholar 

  17. Nakaya T, Fujinaga K, Kishi M, et al. Nonsense mutations in the vpr gene of HIV-1 during in vitro passage and in HIV-1 carrier-derived peripheral blood mononuclear cells. FEBS Letts 1994; 354: 17-22.

    Google Scholar 

  18. Macreadie IG, Arunagiri CK, Hewish DR, White JF, Azad AA. Extracellular addition of a domain of HIV-1 Vpr containing the amino acid sequence motif H(S/F)RIG cause cell membrane permeabilization and death. Mol Microbiol 1996; 19: 1185-1192.

    Google Scholar 

  19. Adachi A, Gendelman HE, Koenig S, et al. Production of acquired immunodeficiency syndrome associated retrovirus in human and nonhuman cells transfected with an infectious molecular clone. J Virol 1986; 59: 284-291.

    Google Scholar 

  20. Smith DB, Johnson KS. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene 1988; 67: 31-40.

    Google Scholar 

  21. Harlow E, Lane D. Antibodies: A Laboratory Manual. New York, NY (USA): Cold Spring Harbor Laboratory Press, 1988: 354-355.

  22. Nicolas P, Mor A. Peptides as weapons against microorganisms in the chemical defense system of vertebrates. Annu Rev Microbiol 1995; 49: 277-304.

    Google Scholar 

  23. Shapiro H. Practical Flow Cytometry. New York, NY (USA): Wiley-Liss, 1994: 251-262.

  24. Gartner F, Voos W, Querol A, et al. Mitochondrial import of subunit Va of cytochrome c oxidase characterised with yeast mutants. J Biol Chem 1995; 270: 3788-3795.

    Google Scholar 

  25. Petit PX, O'Connor JE, Grinwald D, Brown SC. Analysis of the membrane potential of rat-and mouse-liver mitochondria by flow cytometry and possible applications. Eur J Biochem 1990; 194: 389-397.

    Google Scholar 

  26. Zamzami N, Marchetti P, Castedo M, et al. Sequential reduction of mitochondrial transmembrane potential and generation of reactive oxygen species in early programmed cell death. J Exp Med 1995; 182: 367-377

    Google Scholar 

  27. Petit PX, Le Couer H, Zorn E, Daught C, Mignotte B, Gougeon ML. Alteration of mitochondrial structure and function are early events of dexamethasone-induced thymocyte apoptosis. J Cell Biol 1995; 130: 157-167.

    Google Scholar 

  28. Lu YL, Spearman P, Ratner L. Human immunodeficiency virus type 1 viral protein R localization in infected cells and virions. J Virol 1993; 67: 6542-6550.

    Google Scholar 

  29. Gorczyka W, Bruno S, Darzynkiewicz RJ, Gong J, Darzynkiewicy Z. DNA strand breaks during apoptosis. Their early in situ detection by terminal deoxynucleotidyl transferase and nick translation assays and prevention by serine protease inhibitors. Inst J Onc 1992; 1: 639-648.

    Google Scholar 

  30. Cohen GM, Sun ZM, Snowden RT, Dinsdale D, Skilleter DN. Key morphological features of apoptosis may occur in the absence of internucleosomal DNA fragmentation. Biochem J 1992; 286: 331-334.

    Google Scholar 

  31. Dancescu M, Rubio-Trujillo M, Biron G, Bron D, Delespesse G, Sarfarti M. Interleukin 4 protects chronic lymphocytic leukemic B cells from death by apoptosis and up regulates Bcl-2 expression. J Exp Med 1992; 176: 1319-1326.

    Google Scholar 

  32. Razvi ES, Welsh RM. Apoptosis in viral infections. Adv virus Res 1995; 45: 1-60

    Google Scholar 

  33. Darzynkiewicy Z, Bruno S, DelBino G, et al. Features of apoptotic cells measured by flow cytometry. Cytometry 1992; 13: 795-808.

    Google Scholar 

  34. Deckwerth TL, Johnson EM. Temporal analysis of events associated with programmed cell death (apoptosis) of sympathetic neurons deprived of nerve growth factor. J Cell Biol 1993; 123: 1207-1222.

    Google Scholar 

  35. Vayssiere JL, Petit PX, Risler Y, Mignotte B. Commitment to apoptosis is associated with changes in mitochondrial biogenesis and activity in cell lines conditionally immortalised with simian virus 40. Proc Natl Acad Sci USA 1994; 91: 11752-11756.

    Google Scholar 

  36. Zamzami N, Marchetti P, Castedo M, et al. Reduction in mitochondrial potential constitutes an early irreversible step of programmed lymphocyte death in vivo. J Exp Med 1995; 181: 1661-1672.

    Google Scholar 

  37. Castedo M, Macho A, Zamzami N, et al. Mitochondrial perturbations define lymphocytes undergoing apoptotic depletion in vivo. Eur J Immunol 1995; 25: 3277-3284.

    Google Scholar 

  38. Kroemer G, Petit PX, Zamzami N, Vayssiere JL, Mignotte B. The biochemistry of apoptosis. FASEB J 1995; 9: 1277-1287.

    Google Scholar 

  39. Zamzami N, Susin SA, Marchetti P, et al. Mitochondrial control of nuclear apoptosis. J Exp Med 1996; 183: 1533-1544.

    Google Scholar 

  40. Macho A, Castedo M, Marchetti P, et al. Mitochondrial dysfunction in circulating T lymphocytes from human immunodeficiency virus-1 carriers. Blood 1995; 86: 2481-2487.

    Google Scholar 

  41. Somasundaran M, Zapp ML, Beattie LK, et al. Localisation of HIV RNA in mitochondria of infected cells: Potential role in cytopathogenicity. J cell Biol 1994; 126: 1353-1360.

    Google Scholar 

  42. Forte SE, Sullivan JL, Somasundaran M. In vitro characterisation of HIV type 1 biological clones from asymptomatic and symptomatic pediatric patients. AIDS Res Human Retroviruses 1996; 12: 1585-1593.

    Google Scholar 

  43. Levy JA. Pathogenesis of human immunodeficiency virus infection. Microbiol Rev 1993; 57: 183-289.

    Google Scholar 

  44. Meyaard L, Otto SA, Jonker RR, Mijnster MJ, Keet IPM, Miedema F. Programmed death of T cells in HIV-1 infection. Science 1992; 257: 217-219.

    Google Scholar 

  45. Groux H, Torpier G, Monte D, Mouton Y, Capron A, Ameisen JC. Activation-induced death by apoptosis in CD4 T cells from human immunodeficiency virus-infected asymptomatic individuals. J Exp Med 1992; 175: 331-340.

    Google Scholar 

  46. Gougeon ML, Garcia S, Heeney J, et al. Programmed cell death in AIDS-related HIV and SIV infections. AIDS Res Hum Retroviruses 1993; 9: 553-563.

    Google Scholar 

  47. Janossy J, Borthwick N, Lomnitzer R, et al. Lymphocyte activation in HIV-1 infection. I. Predominant proliferative defects among CD45R0+ cells of the CD4 and CD8 lineages. AIDS 1993; 7: 613-624.

    Google Scholar 

  48. Carbonari M, Cibati M, Pesce AM, et al. Frequency of provirusbearing CD4+ cells in HIV type 1 infection correlates with extent of in vitro apoptosis of CD8+ but not of CD4+ cells. AIDS Res Hum Retroviruses 1995; 11: 789-794.

    Google Scholar 

  49. Carbonari M, Cibati M, Cherchi M, et al. Detection and characterization of apoptotic peripheral blood lymphocytes in human immunodeficiency virus infection and cancer chemotherapy by a novel flow immunocytometric method. Blood 1994; 83: 1245-1255.

    Google Scholar 

  50. Re MC, Zauli G, Gibellini D, et al. Uninfected haematopoietic progenitor (CD34+) cells purified from the bone marrow of AIDS patients are committed to apoptotic cell death in culture. AIDS 1993; 7: 1049-1055.

    Google Scholar 

  51. Wei X, Ghosh SK, Taylor ME, et al. Viral dynamics in human immunodeficiency virus type 1 infection. Nature 1995; 273: 117-122.

    Google Scholar 

  52. Ho DD, Neuman AV, Perelson AS, Chen W, Leonard JM, Markowitz M. Rapid turnover of plasma virions and CD4 lymphocytes in HIV-1 infection. Nature 1995; 373: 123-126.

    Google Scholar 

  53. Nardelli B, Gouzaley CJ, Schechter M, Valentine FT. CD4+ blood lymphocytes are rapidly killed in vitro by contact with autologous human immunodeficiency virus-infected cells. Proc Natl Acad Sci USA 1995; 92: 7312-7316.

    Google Scholar 

  54. Pantaleo G, Graziosi C, Demarest JF, et al. HIV infection is active and progressive in lymphoid tissue during the clinically latent stage of the disease. Nature 1993; 362: 355-358.

    Google Scholar 

  55. Embretson J, Zupanuc M, Ribas JL, et al. Massive covert infection of helper T lymphocyte and macrophages by HIV during the incubation period of AIDS. Nature 1993; 362: 359-362.

    Google Scholar 

  56. Banda NK, Bernier J, Kurahara DK, et al. Cross linking of CD4 by human immunodeficiency virus gp120 primes T cells for activation-induced apoptosis. J Exp Med 1992; 176: 1099-1106.

    Google Scholar 

  57. Banda NK, Satterfield WC, Dunlop A, Steimer KS, Kurrie R, Finkel TA. Lack of gp120 induced anergy and apoptosis in chimpanzees is correlated with resistance to AIDS. Apoptosis 1996; 1: 49-62.

    Google Scholar 

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Authors and Affiliations

  1. Division of Biomolecular Engineering, Biomolecular Research Institute, 343 Royal Parade, Parkville, Australia

    C. Arunagiri, I. Macreadie & A. Azad

  2. CSIRO, Division of Biomolecular Engineering, 343 Royal Parade, Parkville, Australia

    D. Hewish

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  1. C. Arunagiri
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  2. I. Macreadie
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  3. D. Hewish
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  4. A. Azad
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Correspondence to A. Azad.

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Arunagiri, C., Macreadie, I., Hewish, D. et al. A C-terminal domain of HIV-1 accessory protein Vpr is involved in penetration, mitochondrial dysfunction and apoptosis of human CD4+ lymphocytes. Apoptosis 2, 69–76 (1997). https://doi.org/10.1023/A:1026487609215

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