Molecular Functional Studies of HIV-1 REV and NEF Proteins

  • Sundararajan Venkatesan
  • Steven M. Holland
  • Nafees Ahmad
  • Paul Wingfield
  • Ratan K. Maitra
  • H. Clifford Lane
Part of the GWUMC Department of Biochemistry Annual Spring Symposia book series (GWUN)


Human immunodeficiency virus type 1 (HIV-1), the etiological agent of AIDS, preferentially infects the CD4+ helper subset of human T lymphocytes. Clinically, HIV-1 infection is characterized by a chronic phase lasting several years with a paucity of infected lymphocytes in the circulation. Notwithstanding, HIV-1 infection of primary human T lymphocytes or CD4+ cell lines in vitro leads to massive acute infection and cell death (20). This dichotomy between the natural history of virus infection and its behavior in tissue culture implies the existence of viral and cellular determinants of viral latency and reactivation.


Human Immunodeficiency Virus Jurkat Cell Provirus Expression Primate Immunodeficiency Virus 
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  1. 1.
    Adachi, A., H. E. Gendelman, S. Koenig, T. Folks, R. Willey, A. Rabson, and M. A. Martin. (1986) Production of acquired immunodeficiency syndrome-associated retrovirus in human and nonhuman cells transfected with an infectious molecular clone. J. Virol. 59, 284–291.PubMedGoogle Scholar
  2. 2.
    Ahmad, N., and Venkatesan, S. (1988) Nef Protein of HIV-1 is a transcriptional repressor of HIV-1 LTR. Science 241, 1481–1485.PubMedCrossRefGoogle Scholar
  3. 3.
    Ahmad, N., Maitra, R. K., and Venkatesan, S. (1989). HIV-1 Rev induced modulation of Nef protein underlies temporal regulation of viral replication. Proc. Natl. Acad. Sci. (USA) 86, 61116115.Google Scholar
  4. 4.
    Allan, J. S., Coligan, J. E., Lee, T. H., McLane, M. F., Kanki, P. J., Groopman, J. E., and Essex, M. (1985) A new HTLV-III/LAV encoded antigen detected by antibodies from AIDS patients. Science 230, 810–813.PubMedCrossRefGoogle Scholar
  5. 5.
    Arya, S.K., Guo, C., Josephs, S.F., and Wong-Staal, F. (1985). Trans-activator gene of human T-lymphotropic virus type III (HTLV-IlI). Science 229, 69–73.PubMedCrossRefGoogle Scholar
  6. 6.
    Bandziulis, R. J., M. S. Swanson, and G. Dreyfuss. (1989) RNA binding proteins as developmental regulators. Genes Develop. 3, 431–437.PubMedCrossRefGoogle Scholar
  7. 7.
    Berkhout, B., Silverman, R., and Jeang, K-T. (1989) Tat trans-activates human immunodeficiency virus through a nascent RNA target. Cell 57, 273–282.CrossRefGoogle Scholar
  8. 8.
    Berkhout, B. R., Gatignol, A., Rabson, A. R., and Jeang, K.-T. (1990) TAR-independent activation of the HIV-1 LTR: Evidence that Tat requires specific regions of the promoter. Cell (In press).Google Scholar
  9. 9.
    Chang, D. D., and P. A. Sharp. (1989) Regulation by HIV Rev depends upon recognition of splice sites. Cell 59, 789–795.Google Scholar
  10. 10.
    Cheng-Mayer, C., Iannello, P., Shaw, K., Luciw, P. A., and Levy, J. A. (1989) Differential effects of nef on HIV replication: Implications for viral pathogenesis in the host. Science 246, 1629–1632.PubMedCrossRefGoogle Scholar
  11. 11.
    Cochrane, A. W., C.-H. Chen, and C. A. Rosen. (1990) Specific interaction of the human immunodeficiency virus Rev protein with a structured region in the env mRNA. Proc. Natl. Acad. Sci. USA 87, 1198–1202.Google Scholar
  12. 12.
    Cullen, B. R. (1986) Trans-activation of human immunodeficiency virus occurs via a bimodal mechanism. Cell 46, 973–81.PubMedCrossRefGoogle Scholar
  13. 13.
    Cullen, B. R., J. Hauber, K. Campbell, J. G. Sodroski, W. A. Haseltine, and C. A. Rosen. (1988) Subcellular localization of the human immunodeficiency virus trans-acting rev gene product. J Virol. 62, 2498–2501.PubMedGoogle Scholar
  14. 14.
    Daly, T. J., Cook, K. S., Gray,G. S., Maione, T. E., and Rusche, J. R. (1989) Specific binding of HIV-1 recombinant Rev protein to the Rev-responsive element in vitro. Nature (London) 342, 816–819.Google Scholar
  15. 15.
    Dayton, A.I., Sodroski, J.G., Rosen, C.A., Goh, W.C., and Haseltine, W.A. (1986). The trans-activator gene of the human T-cell lymphotropic virus type III is required for replication. Cell 44, 941–947.PubMedCrossRefGoogle Scholar
  16. 16.
    Dayton, E. T., Powell,D. M., and Dayton, A. I. (1989) Functional analysis of CAR, the target sequence for the Rev protein of HIV-1. Science 246, 1625–1629.Google Scholar
  17. 17.
    Duh, E. J., Maury, W. J., Folks, T. M., Fauci, A. S., and Rabson, A. B. (1989) Tumor necrosis factor a activates human immunodeficiency virus type 1 through induction of nuclear factor binding to the NF-kB sites in the long terminal repeat. Proc. Natl. Acad. Sci. (USA) 86, 59745978.Google Scholar
  18. 18.
    Emerman, M., R. Vazeur, and K. Peden. (1989) The rev gene product of the human immunodeficiency virus affects envelope-specific RNA localization. Cell 57, 1155–1165.PubMedCrossRefGoogle Scholar
  19. 19.
    Felber, B. K. (1989) Cross-activation of the Rex proteins of HTLV-1 and BLV and of the REV protein of HIV-1 and non-reciprocal interactions with their RNA responsive elements. The New biologist 1, 318–330.PubMedGoogle Scholar
  20. 20.
    Fauci, A. S. (1988) The human immunodeficiency virus: infectivity and mechanisms of pathogenesis. Science 239, 617–622.PubMedCrossRefGoogle Scholar
  21. 21.
    Feinberg, M.B., Jarrett, R.F., Aldovini, A., Gallo, R.C., and Wong-Staal, F. (1986). HTLV-III gene expression and production involve complex regulation at the levels of splicing and translation of RNA. Cell 46, 807–817.PubMedCrossRefGoogle Scholar
  22. 22.
    Fisher, A.G., Feinberg, M.B., Josephs, S.F., Harper, M.E.,Marselle, L.M., Reyes, G., Gonda, M.A., Aldovini, A., Debouk, C., Gallo, R.C., and Wong-Staal, F. (1986). The transactivator gene of HTLV-Ilf is essential for virus replication. Nature 320, 367–371.Google Scholar
  23. 23.
    Franchini, G., Robert-Guroff, M., Ghrayeb, J., Chang, N. T., and Wong-Staal, F. (1986) Cytoplasmic localization of the HTLV III 3’ orf protein in cultured T cells. Virology 155, 593–599.PubMedCrossRefGoogle Scholar
  24. 24.
    Garcia, J. A., Wu, F. K., Mitsuyasu, R., and Gaynor, R. B. (1987) Interactions of cellular proteins involved in the transcriptional regulation of the human immunodeficiency virus. The EMBO J. 6, 3761–3770.Google Scholar
  25. 25.
    Guy, B., Kieny, M.P., Riviere, Y., LePeuch, C., Dott, K., Girard,M., Montagnier, L, and Lecocq, J-P. (1987). HIV F/3’ ORF encodes a phosphorylated GTP-binding protein resembling an oncogene product. Nature 330, 266–269.PubMedCrossRefGoogle Scholar
  26. 26.
    Guy, B., Riviere, Y., Dott, K., Regnault, A., and Kieny, M. P. (1990). Mutational analysis of the HIV-1 nef protein. Virology 176, 413–425.PubMedCrossRefGoogle Scholar
  27. 27.
    Hadzoupoulu-Cladaras, M., Felber,B. K., Cladaras, C., Athanasopoulos, A., Tse,A., and Pavlakis, G. N. (1989) The rev (trs/art) protein of human immunodeficiency virus type I affects viral mRNA and protein expression via cis acting sequences in the env region. J. Virol. 63, 1265 1274.Google Scholar
  28. 28.
    Hammes, S. R., Dixon, E. P., Malim, M. M., Cullen, B. R., and Greene, W. C. (1989). Nef protein of human immunodeficiency virus type 1: Evidence against its role as a transcriptional inhibitor. Proc. Natl. Acad. Sci. (USA) 86, 9549–9553.CrossRefGoogle Scholar
  29. 29.
    Hanly, S. M., L. T. Rimsky,M. H., Malim, J. H. Kim, J. Hauber, M. Duc Dodon, S.-Y. Le, J. V. Maizel, B. R. Cullen, and W. C. Greene. (1989) Comparative analysis of the HTLV-I Rex and HIV-1 Rev trans-regulatory proteins and their RNA response elements. Genes and Develop. 3, 1534–1544.Google Scholar
  30. 30.
    Heaphy, S., C. Dingwall, I. Ernberg, M. J. Gait, S. M. Green, J. Kam, A. D. Lowe, M. Singh, and M. A. Skinner. (1990) HIV-1 regulator of virion expression (Rev) binds to an RNA stem-loop structure located in the Rev response element region. Cell 60, 685–693.Google Scholar
  31. 31.
    Holland, S. M., Ahmad, N., Maitra, R. K., Wingfield, P., and Venkatesan, S. (1990) Human immunodeficiency virus REV protein recognizes a target sequence in the RRE RNA within the context of RNA secondary structure. J. Virol., 64, 5966–5975.PubMedGoogle Scholar
  32. 32.
    Hoxie, J. A., Haggarty, B. S., Rackowski, J. L., Pillsbury, N., Levy, J. A. (1985) Persistent noncytopathic infection of normal human T lymphocytes with AIDS-associated retrovirus. Science 229, 1400–1402.PubMedCrossRefGoogle Scholar
  33. 33.
    Jones, K. A., Kadonaga, J. T., Luciw, P. A., and Tjian, R. (1986) Activation of the AIDS retrovirus promoter by the cellular transcription factor, Spl. Science 232, 755–759.PubMedCrossRefGoogle Scholar
  34. 34.
    Kaminchik, J., Bashan, N., Pinchasi, D., Amit, B., Sarver, N., Johnston, M. I., Fischer, M., Yavin, Z., Gorecki, M., and Panet, A. (1990) Expression and biochemical characterization of human immunodeficiency virus type 1 nef gene product. J. Virol. 64: 3447–3454.PubMedGoogle Scholar
  35. 35.
    Kim, S., Ikeuchi, K., Byrn, R., Groopman, J., and Baltimore, D. (1989). Lack of a negative influence on viral growth by the nef gene of human immunodeficiency virus type 1. Proc. Natl. Acad. Sci. (USA) 86, 9544–9548.CrossRefGoogle Scholar
  36. 36.
    Klatzmann, D., F. Barre-Sinoussi, M. T. Nugeryre, C. Daugnet, E. Vilmer, C. Griscelli, F. BrunVezinet, C. Rouzious, J. C. Gluckman, J. C. Chermann, and L. Montagnier. (1984) Selective trophism of lymphadenopathy associated virus (LAV) for helper-inducer T-lymphocytes. Science 225 59–63.PubMedCrossRefGoogle Scholar
  37. 37.
    Laspia, M. F., Rice, A. P., and Mathews, M. (1989) HIV-1 Tat protein increases transcriptional initiation and stabilizes elongation. Cell 59, 283–292.PubMedCrossRefGoogle Scholar
  38. 38.
    Lazinski, D., E. Grzadzielska, and A. Das. (1989) Sequence-specific recognition of RNA hairpins by bacteriophage antitermination requires a conserved arginine-rich motif. Cell 59, 207–218.PubMedCrossRefGoogle Scholar
  39. 39.
    Luciw, P.A., Cheng-Mayer, C., and Levy, J.A. (1987). Mutational analysis of the human immunodeficiency virus, The orf-B region down-regulates virus replication. Proc. Natl. Acad. Sci. USA 84, 1434–1438.PubMedCrossRefGoogle Scholar
  40. 40.
    Maitra, R. K., Ahmad, N. McCoy, S., Holland, S. M., Lane, H. C., and Venkatesan, S. (1990) Expression of human immunodeficiency virus, type 1 (HIV-1) NEF protein causes repression of HIV-1 provirus expression and LTR transcription in the context of NEF+ cell lines. Manuscript submitted.Google Scholar
  41. 41.
    Malim, M. H., Hauber, J., Le, S.-Y., Maizel,J. V., and Cuilen,B. R. (1989) The HIV-1 rev trans-activator acts through a structured target sequence to activate nuclear export of unspliced viral mRNA. Nature (London) 338, 254–257.Google Scholar
  42. 42.
    Malim, M. H., S. Bohnlein, J. Hauber, and B. R. Cullen. (1989) Functional dissection of the HIV-1 Rev trans-activator; derivation of a trans-dominant repressor of Rev function. Cell 58, 205–214.Google Scholar
  43. 43.
    Malim, M. H., S. Bohnlein, R. Fenrick, S.-Y., Le, J. V. Maizel, and B. R. Cullen. (1989) Functional comparison of the Rev trans-activators encoded by different primate immunodeficiency virus species. Proc. Natl. Acad. Sci. (USA) 6, 8222–8226.Google Scholar
  44. 44.
    Malim, M. H., Tiley,L. S., McCarn, D. F., Rusche, J. R., Hauber,J., and Cullen, B. R. (1990) HIV-1 structural gene expression requires binding of the Rev trans-activator to its RNA target sequence. Cell 60, 675–683.Google Scholar
  45. 45.
    Mattaj, I. W. (1989) A binding consensus: RNA-protein interactions in splicing, snRNPs, and sex. Cell 57, 1–3.PubMedCrossRefGoogle Scholar
  46. 46.
    Muesing, M. A., Smith, D. H., Cabradilla, C. D., Benton, C. V., Lasky, L. A., and Capon, D. J. (1985) Nucleic acid structure and expression of the human AIDS/lymphadenopathy retrovirus. Nature 313, 430–458.CrossRefGoogle Scholar
  47. 47.
    Muesing, M. A., Smith, D. H., and Capon, D. J. (1987) Regulation of mRNA accumulation by a human immunodeficiency virus trans-acivator protein. Cell 48, 691–701.PubMedCrossRefGoogle Scholar
  48. 48.
    Myers, G., Berzofsky, J. A., Rabson, A. B., and Smith. T. F. (1990) Human Retroviruses and AIDS. Los Alamos National Laboratory (Pub.), Los Alamos, New Mexico.Google Scholar
  49. 49.
    Nabel, G., and Baltimore, D. (1987) An inducible expression factor activates expression of human immunodeficiency virus in T cells. Nature 326, 711–713.PubMedCrossRefGoogle Scholar
  50. 50.
    Nebreda, A. R., Bryan, T., Segade, F., Wingfield, P., Venkatesan, S. and Santos. E. (1990) Human immunodeficiency virus type 1 NEF is an unlikely candidate for GTP binding or oncogenic transformation. ( Manuscript submitted).Google Scholar
  51. 51.
    Niederman, T. M. J., Thielan, B. J., and Ratner, L. (1989) Human immunodeficiency virus type 1 negative factor is a transcriptional silencer. Proc. Natl. Acad. Sci. (USA) 86, 1128–1132.CrossRefGoogle Scholar
  52. 52.
    Olsen, H. S., Nelbrook, P., Cochrane, A. W., and Rosen, C. A. (1990) Secondary structure is the major determinant for interaction of HIV rev protein with RNA. Science 247, 845–848.PubMedCrossRefGoogle Scholar
  53. 53.
    Peterlin, B. M., Luciw, P. A., Barr, P. J., and Walker, M. D. (1986) Elevated levels of mRNA can account for the transactivation of human immunodeficiency virus (HIV). Proc. Natl. Acad. Sci. (USA) 83, 9734–9738.CrossRefGoogle Scholar
  54. 54.
    Query, C. C., R. C. Bentley, and J. D. Keene. (1989) An RNA recognition motif identified within a defined Ul RNA binding domain of the 70K Ul snRNP protein. Cell 57, 89–101.PubMedCrossRefGoogle Scholar
  55. 55.
    Rosen, C. A., Sodroski, J. G., and Haseltine, W. A. (1985) Location of cis-acting regulatory sequences in the human T cell lymphotropic virus type III (HTLV-III/LAV) long terminal repeat. Cell 41, 813–823.PubMedCrossRefGoogle Scholar
  56. 56.
    Schwartz, S., Felber, B. K., Benko, D. M., Fenyo, E.-M., and Pavlakis, G. N. (1990) Cloning and functional analysis of multiply spliced mRNA species of HIV-1. J. Virol. 64, 2519–2529.PubMedGoogle Scholar
  57. 57.
    Shaw, J-P., Utz, P. J., Durand, D. B., Toole, J. J., Emmet, E. A., and Crabtree, G. R. (1988) Identification of a putative regulator of early T cell activation genes. Science 241, 202–205PubMedCrossRefGoogle Scholar
  58. 58.
    Siekevitz, M., Josephs, S.F., Dukovich, M., Peffer, N., Wong-Staal, F., Greene, W.C. (1988). Activation of the HIV-1 LTR by T Cell Mitogens and the Trans-Activator Protein of HTLV-1. Science 238, 1575–1578.CrossRefGoogle Scholar
  59. 59.
    Sodroski, J., Goh, W.C., Rosen, C., Dayton, A., Terwilliger, E.,and Haseltine, W.A. (1986). A second post-transcriptional transactivator gene required for HTLV-Ill replication. Nature 321, 412–417.Google Scholar
  60. 60.
    Southgate, C., Zapp, M. L., and Green, M. R. (1990). Activation of transcription by HIV-1 Tat protein tethered to nascent RNA through another protein. Nature 345, 640–642.PubMedCrossRefGoogle Scholar
  61. 61.
    Terwilliger, E., Sodroski, J.G., Rosen, C.A., and Haseltine, W.A. (1986). Effects of mutations within the 3’ orf open reading frame region of human T-cell lymphotropic virus type III (HTLVIII/LAV) on replication and cytopathogenicity. J. Virol. 60, 754–760.PubMedGoogle Scholar
  62. 62.
    Terwilliger, E., Langhoff, E., Obaru, K., Haseltine, W. (1990) Pleiotropic effects upon HIV replication induced by the nef gene. VI International Conference on AIDS. vol. 2, p133.Google Scholar
  63. 63.
    Turner, D. H., N. Sugimoto, J. A. Jaeger, C. A. Longfellow, S. M. Freier, and R. Kierzek. (1987) Improved parameters for prediction of RNA structure. Cold Spring Harbor Symp. Quant. Biol. 52, 123–131.CrossRefGoogle Scholar
  64. 64.
    York-Higgins, D., Cheng-Mayer, C., Bauer, D., Levy, J. A., and Dina, D. (1990) Human Immunodeficiency virus type 1 cellular host range, and cytopathicity are linked to the envelope region of the viral genome. J. Virol. 64, 4016–4020.PubMedGoogle Scholar
  65. 65.
    Zack, J. A., Arrigo, S. J., O’Brien, W. A., and Chen, I. S. Y. (1990) HIV-1 infection of quiescent primary lymphocytes: Molecular analysis reveals a labile, latent viral DNA. Cell 61, 21–222CrossRefGoogle Scholar
  66. 66.
    Zapp, M. L., and M. R. Green. (1989) Sequence-specific RNA binding by the HIV-1 Rev protein. Nature (London) 342, 714–716.CrossRefGoogle Scholar
  67. 67.
    Zuker, M. (1989) Computer prediction of RNA structure. Methods in Enzymol. 180, 262–288.CrossRefGoogle Scholar
  68. 68.
    Zuker, M. (1989) On finding all suboptimal foldings of an RNA molecule. Science 244, 48–52.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1991

Authors and Affiliations

  • Sundararajan Venkatesan
    • 1
  • Steven M. Holland
    • 1
  • Nafees Ahmad
    • 1
  • Paul Wingfield
  • Ratan K. Maitra
    • 1
  • H. Clifford Lane
    • 3
    • 2
  1. 1.Laboratory of Molecular MicrobiologyNational Institute of Allergy and Infectious DiseasesBethesdaUSA
  2. 2.Laboratory of ImmunoregulationNational Institute of Allergy and Infectious DiseasesBethesdaUSA
  3. 3.Protein Expression LaboratoryNational Institutes of HealthBethesdaUSA

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