Human immunodeficiency virus type 1 Nef associates with a cellular serine kinase in T lymphocytes (original) (raw)

Abstract

With T-cell lines constitutively expressing Nef from the SF2 strain of human immunodeficiency virus type 1 (HIV-1SF2) in the form of a hybrid CD8-Nef fusion protein or T-cell lines chronically infected with HIV-1SF2, a cellular serine kinase was found that specifically associates with Nef. Proteins of 62 kDa and 72 kDa, which coimmunoprecipitated with Nef, were phosphorylated in in vitro kinase assays. This Nef-associated serine kinase activity was not blocked by inhibitors of protein kinase C or protein kinase A and was lost when Nef was truncated at amino acid 94 or 99. These findings present evidence that a serine kinase activity is associated with Nef expressed in human T lymphocytes.

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Selected References

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  1. Ahmad N., Venkatesan S. Nef protein of HIV-1 is a transcriptional repressor of HIV-1 LTR. Science. 1988 Sep 16;241(4872):1481–1485. doi: 10.1126/science.3262235. [DOI] [PubMed] [Google Scholar]
  2. Allan J. S., Coligan J. E., Lee T. H., McLane M. F., Kanki P. J., Groopman J. E., Essex M. A new HTLV-III/LAV encoded antigen detected by antibodies from AIDS patients. Science. 1985 Nov 15;230(4727):810–813. doi: 10.1126/science.2997921. [DOI] [PubMed] [Google Scholar]
  3. Backer J. M., Mendola C. E., Fairhurst J. L., Kovesdi I. The HIV-1 nef protein does not have guanine nucleotide binding, GTPase, or autophosphorylating activities. AIDS Res Hum Retroviruses. 1991 Dec;7(12):1015–1020. doi: 10.1089/aid.1991.7.1015. [DOI] [PubMed] [Google Scholar]
  4. Cheng-Mayer C., Iannello P., Shaw K., Luciw P. A., Levy J. A. Differential effects of nef on HIV replication: implications for viral pathogenesis in the host. Science. 1989 Dec 22;246(4937):1629–1632. doi: 10.1126/science.2531920. [DOI] [PubMed] [Google Scholar]
  5. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  6. Colombini S., Arya S. K., Reitz M. S., Jagodzinski L., Beaver B., Wong-Staal F. Structure of simian immunodeficiency virus regulatory genes. Proc Natl Acad Sci U S A. 1989 Jul;86(13):4813–4817. doi: 10.1073/pnas.86.13.4813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cooper J. A., Sefton B. M., Hunter T. Detection and quantification of phosphotyrosine in proteins. Methods Enzymol. 1983;99:387–402. doi: 10.1016/0076-6879(83)99075-4. [DOI] [PubMed] [Google Scholar]
  8. Franchini G., Robert-Guroff M., Ghrayeb J., Chang N. T., Wong-Staal F. Cytoplasmic localization of the HTLV-III 3' orf protein in cultured T cells. Virology. 1986 Dec;155(2):593–599. doi: 10.1016/0042-6822(86)90219-9. [DOI] [PubMed] [Google Scholar]
  9. Garcia J. V., Alfano J., Miller A. D. The negative effect of human immunodeficiency virus type 1 Nef on cell surface CD4 expression is not species specific and requires the cytoplasmic domain of CD4. J Virol. 1993 Mar;67(3):1511–1516. doi: 10.1128/jvi.67.3.1511-1516.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Garcia J. V., Miller A. D. Serine phosphorylation-independent downregulation of cell-surface CD4 by nef. Nature. 1991 Apr 11;350(6318):508–511. doi: 10.1038/350508a0. [DOI] [PubMed] [Google Scholar]
  11. Guy B., Kieny M. P., Riviere Y., Le Peuch C., Dott K., Girard M., Montagnier L., Lecocq J. P. HIV F/3' orf encodes a phosphorylated GTP-binding protein resembling an oncogene product. Nature. 1987 Nov 19;330(6145):266–269. doi: 10.1038/330266a0. [DOI] [PubMed] [Google Scholar]
  12. Guy B., Rivière Y., Dott K., Regnault A., Kieny M. P. Mutational analysis of the HIV nef protein. Virology. 1990 Jun;176(2):413–425. doi: 10.1016/0042-6822(90)90011-f. [DOI] [PubMed] [Google Scholar]
  13. Hammes S. R., Dixon E. P., Malim M. H., Cullen B. R., Greene W. C. Nef protein of human immunodeficiency virus type 1: evidence against its role as a transcriptional inhibitor. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9549–9553. doi: 10.1073/pnas.86.23.9549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Harris M., Coates K. Identification of cellular proteins that bind to the human immunodeficiency virus type 1 nef gene product in vitro: a role for myristylation. J Gen Virol. 1993 Aug;74(Pt 8):1581–1589. doi: 10.1099/0022-1317-74-8-1581. [DOI] [PubMed] [Google Scholar]
  15. Harris M., Hislop S., Patsilinacos P., Neil J. C. In vivo derived HIV-1 nef gene products are heterogeneous and lack detectable nucleotide binding activity. AIDS Res Hum Retroviruses. 1992 May;8(5):537–543. doi: 10.1089/aid.1992.8.537. [DOI] [PubMed] [Google Scholar]
  16. Herrmann C. H., Su L. K., Harlow E. Adenovirus E1A is associated with a serine/threonine protein kinase. J Virol. 1991 Nov;65(11):5848–5859. doi: 10.1128/jvi.65.11.5848-5859.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kaminchik J., Bashan N., Pinchasi D., Amit B., Sarver N., Johnston M. I., Fischer M., Yavin Z., Gorecki M., Panet A. Expression and biochemical characterization of human immunodeficiency virus type 1 nef gene product. J Virol. 1990 Jul;64(7):3447–3454. doi: 10.1128/jvi.64.7.3447-3454.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kaminsky L. S., McHugh T., Stites D., Volberding P., Henle G., Henle W., Levy J. A. High prevalence of antibodies to acquired immune deficiency syndrome (AIDS)-associated retrovirus (ARV) in AIDS and related conditions but not in other disease states. Proc Natl Acad Sci U S A. 1985 Aug;82(16):5535–5539. doi: 10.1073/pnas.82.16.5535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kestler H. W., 3rd, Ringler D. J., Mori K., Panicali D. L., Sehgal P. K., Daniel M. D., Desrosiers R. C. Importance of the nef gene for maintenance of high virus loads and for development of AIDS. Cell. 1991 May 17;65(4):651–662. doi: 10.1016/0092-8674(91)90097-i. [DOI] [PubMed] [Google Scholar]
  20. Kim S., Ikeuchi K., Byrn R., Groopman J., Baltimore D. Lack of a negative influence on viral growth by the nef gene of human immunodeficiency virus type 1. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9544–9548. doi: 10.1073/pnas.86.23.9544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Luciw P. A., Cheng-Mayer C., Levy J. A. Mutational analysis of the human immunodeficiency virus: the orf-B region down-regulates virus replication. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1434–1438. doi: 10.1073/pnas.84.5.1434. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Luria S., Chambers I., Berg P. Expression of the type 1 human immunodeficiency virus Nef protein in T cells prevents antigen receptor-mediated induction of interleukin 2 mRNA. Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5326–5330. doi: 10.1073/pnas.88.12.5326. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Maitra R. K., Ahmad N., Holland S. M., Venkatesan S. Human immunodeficiency virus type 1 (HIV-1) provirus expression and LTR transcription are repressed in NEF-expressing cell lines. Virology. 1991 Jun;182(2):522–533. doi: 10.1016/0042-6822(91)90593-z. [DOI] [PubMed] [Google Scholar]
  24. Mariani R., Skowronski J. CD4 down-regulation by nef alleles isolated from human immunodeficiency virus type 1-infected individuals. Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5549–5553. doi: 10.1073/pnas.90.12.5549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Matsuura Y., Maekawa M., Hattori S., Ikegami N., Hayashi A., Yamazaki S., Morita C., Takebe Y. Purification and characterization of human immunodeficiency virus type 1 nef gene product expressed by a recombinant baculovirus. Virology. 1991 Oct;184(2):580–586. doi: 10.1016/0042-6822(91)90428-e. [DOI] [PubMed] [Google Scholar]
  26. Nebreda A. R., Bryan T., Segade F., Wingfield P., Venkatesan S., Santos E. Biochemical and biological comparison of HIV-1 NEF and ras gene products. Virology. 1991 Jul;183(1):151–159. doi: 10.1016/0042-6822(91)90128-x. [DOI] [PubMed] [Google Scholar]
  27. Niederman T. M., Garcia J. V., Hastings W. R., Luria S., Ratner L. Human immunodeficiency virus type 1 Nef protein inhibits NF-kappa B induction in human T cells. J Virol. 1992 Oct;66(10):6213–6219. doi: 10.1128/jvi.66.10.6213-6219.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Niederman T. M., Hastings W. R., Luria S., Bandres J. C., Ratner L. HIV-1 Nef protein inhibits the recruitment of AP-1 DNA-binding activity in human T-cells. Virology. 1993 May;194(1):338–344. doi: 10.1006/viro.1993.1264. [DOI] [PubMed] [Google Scholar]
  29. Niederman T. M., Thielan B. J., Ratner L. Human immunodeficiency virus type 1 negative factor is a transcriptional silencer. Proc Natl Acad Sci U S A. 1989 Feb;86(4):1128–1132. doi: 10.1073/pnas.86.4.1128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Poulin L., Levy J. A. The HIV-1 nef gene product is associated with phosphorylation of a 46 kD cellular protein. AIDS. 1992 Aug;6(8):787–791. doi: 10.1097/00002030-199208000-00003. [DOI] [PubMed] [Google Scholar]
  31. Sawai E. T., Butel J. S. Association of a cellular heat shock protein with simian virus 40 large T antigen in transformed cells. J Virol. 1989 Sep;63(9):3961–3973. doi: 10.1128/jvi.63.9.3961-3973.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schwartz O., Rivière Y., Heard J. M., Danos O. Reduced cell surface expression of processed human immunodeficiency virus type 1 envelope glycoprotein in the presence of Nef. J Virol. 1993 Jun;67(6):3274–3280. doi: 10.1128/jvi.67.6.3274-3280.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Shibata R., Miura T., Hayami M., Ogawa K., Sakai H., Kiyomasu T., Ishimoto A., Adachi A. Mutational analysis of the human immunodeficiency virus type 2 (HIV-2) genome in relation to HIV-1 and simian immunodeficiency virus SIV (AGM). J Virol. 1990 Feb;64(2):742–747. doi: 10.1128/jvi.64.2.742-747.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Skowronski J., Parks D., Mariani R. Altered T cell activation and development in transgenic mice expressing the HIV-1 nef gene. EMBO J. 1993 Feb;12(2):703–713. doi: 10.1002/j.1460-2075.1993.tb05704.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Terwilliger E., Sodroski J. G., Rosen C. A., Haseltine W. A. Effects of mutations within the 3' orf open reading frame region of human T-cell lymphotropic virus type III (HTLV-III/LAV) on replication and cytopathogenicity. J Virol. 1986 Nov;60(2):754–760. doi: 10.1128/jvi.60.2.754-760.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Tsunetsugu-Yokota Y., Matsuda S., Maekawa M., Saito T., Takemori T., Takebe Y. Constitutive expression of the nef gene suppresses human immunodeficiency virus type 1 (HIV-1) replication in monocytic cell lines. Virology. 1992 Dec;191(2):960–963. doi: 10.1016/0042-6822(92)90272-q. [DOI] [PubMed] [Google Scholar]
  37. Wolber V., Rensland H., Brandmeier B., Sagemann M., Hoffmann R., Kalbitzer H. R., Wittinghofer A. Expression, purification and biochemical characterisation of the human immunodeficiency virus 1 nef gene product. Eur J Biochem. 1992 May 1;205(3):1115–1121. doi: 10.1111/j.1432-1033.1992.tb16880.x. [DOI] [PubMed] [Google Scholar]
  38. Yu G., Felsted R. L. Effect of myristoylation on p27 nef subcellular distribution and suppression of HIV-LTR transcription. Virology. 1992 Mar;187(1):46–55. doi: 10.1016/0042-6822(92)90293-x. [DOI] [PubMed] [Google Scholar]
  39. Zazopoulos E., Haseltine W. A. Mutational analysis of the human immunodeficiency virus type 1 Eli Nef function. Proc Natl Acad Sci U S A. 1992 Jul 15;89(14):6634–6638. doi: 10.1073/pnas.89.14.6634. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. de Ronde A., Klaver B., Keulen W., Smit L., Goudsmit J. Natural HIV-1 NEF accelerates virus replication in primary human lymphocytes. Virology. 1992 May;188(1):391–395. doi: 10.1016/0042-6822(92)90772-h. [DOI] [PubMed] [Google Scholar]