Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1996 Sep 3;93(18):9571–9576. doi: 10.1073/pnas.93.18.9571

Apoptosis mediated by HIV protease is preceded by cleavage of Bcl-2.

P R Strack 1, M W Frey 1, C J Rizzo 1, B Cordova 1, H J George 1, R Meade 1, S P Ho 1, J Corman 1, R Tritch 1, B D Korant 1
PMCID: PMC38469  PMID: 8790371

Abstract

Expression of the human immunodeficiency virus type 1 (HIV) protease in cultured cells leads to apoptosis, preceded by cleavage of bcl-2, a key negative regulator of cell death. In contrast, a high level of bcl-2 protects cells in vitro and in vivo from the viral protease and prevents cell death following HIV infection of human lymphocytes, while reducing the yields of viral structural proteins, infectivity, and tumor necrosis factor alpha. We present a model for HIV replication in which the viral protease depletes the infected cells of bcl-2, leading to oxidative stress-dependent activation of NF kappa B, a cellular factor required for HIV transcription, and ultimately to cell death. Purified bcl-2 is cleaved by HIV protease between phenylalanine 112 and alanine 113. The results suggest a new option for HIV gene therapy; bcl-2 muteins that have noncleavable alterations surrounding the HIV protease cleavage site.

Full text

PDF
9572

Images in this article

9572Fig. 1
on p.9572
9573Fig. 2
on p.9573
9574Fig. 3
on p.9574

Selected References

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

  1. Alnemri E. S., Robertson N. M., Fernandes T. F., Croce C. M., Litwack G. Overexpressed full-length human BCL2 extends the survival of baculovirus-infected Sf9 insect cells. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7295–7299. doi: 10.1073/pnas.89.16.7295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ameisen J. C. Programmed cell death (apoptosis) and cell survival regulation: relevance to AIDS and cancer. AIDS. 1994 Sep;8(9):1197–1213. doi: 10.1097/00002030-199409000-00001. [DOI] [PubMed] [Google Scholar]
  3. Antoni B. A., Sabbatini P., Rabson A. B., White E. Inhibition of apoptosis in human immunodeficiency virus-infected cells enhances virus production and facilitates persistent infection. J Virol. 1995 Apr;69(4):2384–2392. doi: 10.1128/jvi.69.4.2384-2392.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Biswas D. K., Dezube B. J., Ahlers C. M., Pardee A. B. Pentoxifylline inhibits HIV-1 LTR-driven gene expression by blocking NF-kappa B action. J Acquir Immune Defic Syndr. 1993 Jul;6(7):778–786. [PubMed] [Google Scholar]
  5. Carson W. E., Haldar S., Baiocchi R. A., Croce C. M., Caligiuri M. A. The c-kit ligand suppresses apoptosis of human natural killer cells through the upregulation of bcl-2. Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7553–7557. doi: 10.1073/pnas.91.16.7553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. De Rossi A., Ometto L., Roncella S., D'Andrea E., Menin C., Calderazzo F., Rowe M., Ferrarini M., Chieco-Bianchi L. HIV-1 induces down-regulation of bcl-2 expression and death by apoptosis of EBV-immortalized B cells: a model for a persistent "self-limiting" HIV-1 infection. Virology. 1994 Jan;198(1):234–244. doi: 10.1006/viro.1994.1026. [DOI] [PubMed] [Google Scholar]
  7. Esterbauer H., Cheeseman K. H. Determination of aldehydic lipid peroxidation products: malonaldehyde and 4-hydroxynonenal. Methods Enzymol. 1990;186:407–421. doi: 10.1016/0076-6879(90)86134-h. [DOI] [PubMed] [Google Scholar]
  8. Freitas J. P., Filipe P. M. Pentoxifylline. A hydroxyl radical scavenger. Biol Trace Elem Res. 1995 Jan-Mar;47(1-3):307–311. doi: 10.1007/BF02790131. [DOI] [PubMed] [Google Scholar]
  9. Gougeon M. L., Montagnier L. Apoptosis in AIDS. Science. 1993 May 28;260(5112):1269–1270. doi: 10.1126/science.8098552. [DOI] [PubMed] [Google Scholar]
  10. Kane D. J., Sarafian T. A., Anton R., Hahn H., Gralla E. B., Valentine J. S., Ord T., Bredesen D. E. Bcl-2 inhibition of neural death: decreased generation of reactive oxygen species. Science. 1993 Nov 19;262(5137):1274–1277. doi: 10.1126/science.8235659. [DOI] [PubMed] [Google Scholar]
  11. Kaplan A. H., Swanstrom R. Human immunodeficiency virus type 1 Gag proteins are processed in two cellular compartments. Proc Natl Acad Sci U S A. 1991 May 15;88(10):4528–4532. doi: 10.1073/pnas.88.10.4528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kerr J. F., Wyllie A. H., Currie A. R. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer. 1972 Aug;26(4):239–257. doi: 10.1038/bjc.1972.33. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Korant B. D., Rizzo C. J. An E. coli expression system which detoxifies the HIV protease. Biomed Biochim Acta. 1991;50(4-6):643–646. [PubMed] [Google Scholar]
  14. Kornbluth R. S. Significance of T cell apoptosis for macrophages in HIV infection. J Leukoc Biol. 1994 Sep;56(3):247–256. doi: 10.1002/jlb.56.3.247. [DOI] [PubMed] [Google Scholar]
  15. Korsmeyer S. J. Regulators of cell death. Trends Genet. 1995 Mar;11(3):101–105. doi: 10.1016/S0168-9525(00)89010-1. [DOI] [PubMed] [Google Scholar]
  16. Kräusslich H. G. Human immunodeficiency virus proteinase dimer as component of the viral polyprotein prevents particle assembly and viral infectivity. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3213–3217. doi: 10.1073/pnas.88.8.3213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lam P. Y., Jadhav P. K., Eyermann C. J., Hodge C. N., Ru Y., Bacheler L. T., Meek J. L., Otto M. J., Rayner M. M., Wong Y. N. Rational design of potent, bioavailable, nonpeptide cyclic ureas as HIV protease inhibitors. Science. 1994 Jan 21;263(5145):380–384. doi: 10.1126/science.8278812. [DOI] [PubMed] [Google Scholar]
  18. Levine B., Huang Q., Isaacs J. T., Reed J. C., Griffin D. E., Hardwick J. M. Conversion of lytic to persistent alphavirus infection by the bcl-2 cellular oncogene. Nature. 1993 Feb 25;361(6414):739–742. doi: 10.1038/361739a0. [DOI] [PubMed] [Google Scholar]
  19. Levy Y., Brouet J. C. Interleukin-10 prevents spontaneous death of germinal center B cells by induction of the bcl-2 protein. J Clin Invest. 1994 Jan;93(1):424–428. doi: 10.1172/JCI116977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lin K. I., Lee S. H., Narayanan R., Baraban J. M., Hardwick J. M., Ratan R. R. Thiol agents and Bcl-2 identify an alphavirus-induced apoptotic pathway that requires activation of the transcription factor NF-kappa B. J Cell Biol. 1995 Dec;131(5):1149–1161. doi: 10.1083/jcb.131.5.1149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Memon S. A., Petrak D., Moreno M. B., Zacharchuk C. M. A simple assay for examining the effect of transiently expressed genes on programmed cell death. J Immunol Methods. 1995 Mar 13;180(1):15–24. doi: 10.1016/0022-1759(94)00294-7. [DOI] [PubMed] [Google Scholar]
  22. Meyaard L., Otto S. A., Jonker R. R., Mijnster M. J., Keet R. P., Miedema F. Programmed death of T cells in HIV-1 infection. Science. 1992 Jul 10;257(5067):217–219. doi: 10.1126/science.1352911. [DOI] [PubMed] [Google Scholar]
  23. Nakayama K., Nakayama K., Negishi I., Kuida K., Shinkai Y., Louie M. C., Fields L. E., Lucas P. J., Stewart V., Alt F. W. Disappearance of the lymphoid system in Bcl-2 homozygous mutant chimeric mice. Science. 1993 Sep 17;261(5128):1584–1588. doi: 10.1126/science.8372353. [DOI] [PubMed] [Google Scholar]
  24. Olsen C. W., Kehren J. C., Dybdahl-Sissoko N. R., Hinshaw V. S. bcl-2 alters influenza virus yield, spread, and hemagglutinin glycosylation. J Virol. 1996 Jan;70(1):663–666. doi: 10.1128/jvi.70.1.663-666.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Poorman R. A., Tomasselli A. G., Heinrikson R. L., Kézdy F. J. A cumulative specificity model for proteases from human immunodeficiency virus types 1 and 2, inferred from statistical analysis of an extended substrate data base. J Biol Chem. 1991 Aug 5;266(22):14554–14561. [PubMed] [Google Scholar]
  26. Reed J. C., Stein C., Subasinghe C., Haldar S., Croce C. M., Yum S., Cohen J. Antisense-mediated inhibition of BCL2 protooncogene expression and leukemic cell growth and survival: comparisons of phosphodiester and phosphorothioate oligodeoxynucleotides. Cancer Res. 1990 Oct 15;50(20):6565–6570. [PubMed] [Google Scholar]
  27. Rivière Y., Blank V., Kourilsky P., Israël A. Processing of the precursor of NF-kappa B by the HIV-1 protease during acute infection. Nature. 1991 Apr 18;350(6319):625–626. doi: 10.1038/350625a0. [DOI] [PubMed] [Google Scholar]
  28. Rizzo C. J., Korant B. D. Genetic approaches designed to minimize cytotoxicity of retroviral protease. Methods Enzymol. 1994;241:16–29. doi: 10.1016/0076-6879(94)41056-9. [DOI] [PubMed] [Google Scholar]
  29. Schreck R., Rieber P., Baeuerle P. A. Reactive oxygen intermediates as apparently widely used messengers in the activation of the NF-kappa B transcription factor and HIV-1. EMBO J. 1991 Aug;10(8):2247–2258. doi: 10.1002/j.1460-2075.1991.tb07761.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Staal F. J., Roederer M., Herzenberg L. A., Herzenberg L. A. Intracellular thiols regulate activation of nuclear factor kappa B and transcription of human immunodeficiency virus. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9943–9947. doi: 10.1073/pnas.87.24.9943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Subramanian T., Boyd J. M., Chinnadurai G. Functional substitution identifies a cell survival promoting domain common to adenovirus E1B 19 kDa and Bcl-2 proteins. Oncogene. 1995 Dec 7;11(11):2403–2409. [PubMed] [Google Scholar]
  32. Terai C., Kornbluth R. S., Pauza C. D., Richman D. D., Carson D. A. Apoptosis as a mechanism of cell death in cultured T lymphoblasts acutely infected with HIV-1. J Clin Invest. 1991 May;87(5):1710–1715. doi: 10.1172/JCI115188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Tomasselli A. G., Heinrikson R. L. Specificity of retroviral proteases: an analysis of viral and nonviral protein substrates. Methods Enzymol. 1994;241:279–301. doi: 10.1016/0076-6879(94)41069-0. [DOI] [PubMed] [Google Scholar]
  34. Vlasuk G. P., Waxman L., Davis L. J., Dixon R. A., Schultz L. D., Hofmann K. J., Tung J. S., Schulman C. A., Ellis R. W., Bencen G. H. Purification and characterization of human immunodeficiency virus (HIV) core precursor (p55) expressed in Saccharomyces cerevisiae. J Biol Chem. 1989 Jul 15;264(20):12106–12112. [PubMed] [Google Scholar]
  35. Weissman D., Poli G., Fauci A. S. Interleukin 10 blocks HIV replication in macrophages by inhibiting the autocrine loop of tumor necrosis factor alpha and interleukin 6 induction of virus. AIDS Res Hum Retroviruses. 1994 Oct;10(10):1199–1206. doi: 10.1089/aid.1994.10.1199. [DOI] [PubMed] [Google Scholar]
  36. Westendorp M. O., Frank R., Ochsenbauer C., Stricker K., Dhein J., Walczak H., Debatin K. M., Krammer P. H. Sensitization of T cells to CD95-mediated apoptosis by HIV-1 Tat and gp120. Nature. 1995 Jun 8;375(6531):497–500. doi: 10.1038/375497a0. [DOI] [PubMed] [Google Scholar]
  37. Wolff J. A., Malone R. W., Williams P., Chong W., Acsadi G., Jani A., Felgner P. L. Direct gene transfer into mouse muscle in vivo. Science. 1990 Mar 23;247(4949 Pt 1):1465–1468. doi: 10.1126/science.1690918. [DOI] [PubMed] [Google Scholar]
  38. Zhang D., Zhang N., Wick M. M., Byrn R. A. HIV type 1 protease activation of NF-kappa B within T lymphoid cells. AIDS Res Hum Retroviruses. 1995 Feb;11(2):223–230. doi: 10.1089/aid.1995.11.223. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES