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J Virol. 1994 May; 68(5): 3343–3353.
PMCID: PMC236825

Effects of the Tat basic domain on human immunodeficiency virus type 1 transactivation, using chemically synthesized Tat protein and Tat peptides.


To study the structure relationship of different Tat domains, the full-length Tat protein Tat1-86, the gene product of the first exon Tat1-72 which retains full activity of the protein, and a panel of shorter peptides mimicking different regions of the primary structure of the Tat protein were chemically synthesized by the solid-phase method, using an efficient protocol. Synthetic Tat1-86 and Tat1-72 transactivated beta-galactosidase activity in HeLa cells containing the lacZ gene under the control of the human immunodeficiency virus type 1 long terminal repeat. Analyses of the activity of Tat1-86 and Tat1-72 with the sulfhydryl of cysteine residues free or protected by the acetamidomethyl group showed that only the Tat fragments with deprotected cysteine residues retain transactivation ability. In contrast, peptide Tat1-48 was inactive, with cysteine residues either free or protected. Similarly, other shorter synthetic peptides covering the different Tat domains were inactive. Interestingly, when peptides Tat1-48 and Tat38-60 were used simultaneously, a significant transactivation was obtained. This result suggests that both peptide domains are implicated in transactivation, probably by acting at two different sites. This permits us to propose a fundamentally new step in the understanding of the molecular mechanism of Tat transactivation.

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  • Anfinsen CB. Principles that govern the folding of protein chains. Science. 1973 Jul 20;181(4096):223–230. [PubMed]
  • Arya SK, Guo C, Josephs SF, Wong-Staal F. Trans-activator gene of human T-lymphotropic virus type III (HTLV-III). Science. 1985 Jul 5;229(4708):69–73. [PubMed]
  • Barry PA, Pratt-Lowe E, Unger RE, Luciw PA. Cellular factors regulate transactivation of human immunodeficiency virus type 1. J Virol. 1991 Mar;65(3):1392–1399. [PMC free article] [PubMed]
  • Braddock M, Chambers A, Wilson W, Esnouf MP, Adams SE, Kingsman AJ, Kingsman SM. HIV-1 TAT "activates" presynthesized RNA in the nucleus. Cell. 1989 Jul 28;58(2):269–279. [PubMed]
  • Charneau P, Alizon M, Clavel F. A second origin of DNA plus-strand synthesis is required for optimal human immunodeficiency virus replication. J Virol. 1992 May;66(5):2814–2820. [PMC free article] [PubMed]
  • Chavez LG, Jr, Scheraga HA. Intrinsic stabilities of portions of the ribonuclease molecule. Biochemistry. 1980 Mar 4;19(5):1005–1012. [PubMed]
  • Chun R, Glabe CG, Fan H. Chemical synthesis of biologically active tat trans-activating protein of human immunodeficiency virus type 1. J Virol. 1990 Jun;64(6):3074–3077. [PMC free article] [PubMed]
  • Cordingley MG, LaFemina RL, Callahan PL, Condra JH, Sardana VV, Graham DJ, Nguyen TM, LeGrow K, Gotlib L, Schlabach AJ, et al. Sequence-specific interaction of Tat protein and Tat peptides with the transactivation-responsive sequence element of human immunodeficiency virus type 1 in vitro. Proc Natl Acad Sci U S A. 1990 Nov;87(22):8985–8989. [PMC free article] [PubMed]
  • Cullen BR. The HIV-1 Tat protein: an RNA sequence-specific processivity factor? Cell. 1990 Nov 16;63(4):655–657. [PubMed]
  • Desai K, Loewenstein PM, Green M. Isolation of a cellular protein that binds to the human immunodeficiency virus Tat protein and can potentiate transactivation of the viral promoter. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):8875–8879. [PMC free article] [PubMed]
  • Dingwall C, Ernberg I, Gait MJ, Green SM, Heaphy S, Karn J, Lowe AD, Singh M, Skinner MA, Valerio R. Human immunodeficiency virus 1 tat protein binds trans-activation-responsive region (TAR) RNA in vitro. Proc Natl Acad Sci U S A. 1989 Sep;86(18):6925–6929. [PMC free article] [PubMed]
  • Frankel AD, Biancalana S, Hudson D. Activity of synthetic peptides from the Tat protein of human immunodeficiency virus type 1. Proc Natl Acad Sci U S A. 1989 Oct;86(19):7397–7401. [PMC free article] [PubMed]
  • Frankel AD, Bredt DS, Pabo CO. Tat protein from human immunodeficiency virus forms a metal-linked dimer. Science. 1988 Apr 1;240(4848):70–73. [PubMed]
  • Frankel AD, Pabo CO. Cellular uptake of the tat protein from human immunodeficiency virus. Cell. 1988 Dec 23;55(6):1189–1193. [PubMed]
  • Furie B, Schechter AN, Sachs DH, Anfinsen CB. An immunological approach to the conformational equilibrium of staphylococcal nuclease. J Mol Biol. 1975 Mar 15;92(4):497–506. [PubMed]
  • Gatignol A, Buckler-White A, Berkhout B, Jeang KT. Characterization of a human TAR RNA-binding protein that activates the HIV-1 LTR. Science. 1991 Mar 29;251(5001):1597–1600. [PubMed]
  • Gatignol A, Kumar A, Rabson A, Jeang KT. Identification of cellular proteins that bind to the human immunodeficiency virus type 1 trans-activation-responsive TAR element RNA. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7828–7832. [PMC free article] [PubMed]
  • Gaynor R, Soultanakis E, Kuwabara M, Garcia J, Sigman DS. Specific binding of a HeLa cell nuclear protein to RNA sequences in the human immunodeficiency virus transactivating region. Proc Natl Acad Sci U S A. 1989 Jul;86(13):4858–4862. [PMC free article] [PubMed]
  • Green M, Loewenstein PM. Autonomous functional domains of chemically synthesized human immunodeficiency virus tat trans-activator protein. Cell. 1988 Dec 23;55(6):1179–1188. [PubMed]
  • Hauber J, Malim MH, Cullen BR. Mutational analysis of the conserved basic domain of human immunodeficiency virus tat protein. J Virol. 1989 Mar;63(3):1181–1187. [PMC free article] [PubMed]
  • Hurrell JG, Smith JA, Leach SJ. Immunological measurements of conformational motility in regions of the myoglobin molecule. Biochemistry. 1977 Jan 25;16(2):175–185. [PubMed]
  • Jeyapaul J, Reddy MR, Khan SA. Activity of synthetic tat peptides in human immunodeficiency virus type 1 long terminal repeat-promoted transcription in a cell-free system. Proc Natl Acad Sci U S A. 1990 Sep;87(18):7030–7034. [PMC free article] [PubMed]
  • Jeyapaul J, Seshamma T, Khan SA. Synthetic HIV-1 Tat can dissociate HeLa nuclear protein-TAR RNA complexes in vitro: a novel Tat-nuclear protein interaction. Oncogene. 1991 Sep;6(9):1507–1513. [PubMed]
  • Kamine J, Chinnadurai G. Synergistic activation of the human immunodeficiency virus type 1 promoter by the viral Tat protein and cellular transcription factor Sp1. J Virol. 1992 Jun;66(6):3932–3936. [PMC free article] [PubMed]
  • Kato H, Sumimoto H, Pognonec P, Chen CH, Rosen CA, Roeder RG. HIV-1 Tat acts as a processivity factor in vitro in conjunction with cellular elongation factors. Genes Dev. 1992 Apr;6(4):655–666. [PubMed]
  • Krippl B, Ferguson B, Jones N, Rosenberg M, Westphal H. Mapping of functional domains in adenovirus E1A proteins. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7480–7484. [PMC free article] [PubMed]
  • Mann DA, Frankel AD. Endocytosis and targeting of exogenous HIV-1 Tat protein. EMBO J. 1991 Jul;10(7):1733–1739. [PMC free article] [PubMed]
  • Marciniak RA, Garcia-Blanco MA, Sharp PA. Identification and characterization of a HeLa nuclear protein that specifically binds to the trans-activation-response (TAR) element of human immunodeficiency virus. Proc Natl Acad Sci U S A. 1990 May;87(9):3624–3628. [PMC free article] [PubMed]
  • Melton DA, Krieg PA, Rebagliati MR, Maniatis T, Zinn K, Green MR. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. [PMC free article] [PubMed]
  • Nagy JA, Meinwald YC, Scheraga HA. Immunochemical determination of conformational equilibria for fragments of the A alpha chain of fibrinogen. Biochemistry. 1982 Apr 13;21(8):1794–1806. [PubMed]
  • Rappaport J, Lee SJ, Khalili K, Wong-Staal F. The acidic amino-terminal region of the HIV-1 Tat protein constitutes an essential activating domain. New Biol. 1989 Oct;1(1):101–110. [PubMed]
  • Rice AP, Carlotti F. Mutational analysis of the conserved cysteine-rich region of the human immunodeficiency virus type 1 Tat protein. J Virol. 1990 Apr;64(4):1864–1868. [PMC free article] [PubMed]
  • Rice AP, Chan F. Tat protein of human immunodeficiency virus type 1 is a monomer when expressed in mammalian cells. Virology. 1991 Nov;185(1):451–454. [PubMed]
  • Rocancourt D, Bonnerot C, Jouin H, Emerman M, Nicolas JF. Activation of a beta-galactosidase recombinant provirus: application to titration of human immunodeficiency virus (HIV) and HIV-infected cells. J Virol. 1990 Jun;64(6):2660–2668. [PMC free article] [PubMed]
  • Rosen CA, Sodroski JG, Haseltine WA. The location of cis-acting regulatory sequences in the human T cell lymphotropic virus type III (HTLV-III/LAV) long terminal repeat. Cell. 1985 Jul;41(3):813–823. [PubMed]
  • Rosen CA, Terwilliger E, Dayton A, Sodroski JG, Haseltine WA. Intragenic cis-acting art gene-responsive sequences of the human immunodeficiency virus. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2071–2075. [PMC free article] [PubMed]
  • Ruben S, Perkins A, Purcell R, Joung K, Sia R, Burghoff R, Haseltine WA, Rosen CA. Structural and functional characterization of human immunodeficiency virus tat protein. J Virol. 1989 Jan;63(1):1–8. [PMC free article] [PubMed]
  • Sachs DH, Schechter AN, Eastlake A, Anfinsen CB. Antibodies to a distinct antigenic determinant of staphylococcal nuclease. J Immunol. 1972 Dec;109(6):1300–1310. [PubMed]
  • Seigel LJ, Ratner L, Josephs SF, Derse D, Feinberg MB, Reyes GR, O'Brien SJ, Wong-Staal F. Transactivation induced by human T-lymphotropic virus type III (HTLV III) maps to a viral sequence encoding 58 amino acids and lacks tissue specificity. Virology. 1986 Jan 15;148(1):226–231. [PubMed]
  • Sheline CT, Milocco LH, Jones KA. Two distinct nuclear transcription factors recognize loop and bulge residues of the HIV-1 TAR RNA hairpin. Genes Dev. 1991 Dec;5(12B):2508–2520. [PubMed]
  • Siomi H, Shida H, Maki M, Hatanaka M. Effects of a highly basic region of human immunodeficiency virus Tat protein on nucleolar localization. J Virol. 1990 Apr;64(4):1803–1807. [PMC free article] [PubMed]
  • Sodroski J, Rosen C, Wong-Staal F, Salahuddin SZ, Popovic M, Arya S, Gallo RC, Haseltine WA. Trans-acting transcriptional regulation of human T-cell leukemia virus type III long terminal repeat. Science. 1985 Jan 11;227(4683):171–173. [PubMed]
  • Subramanian T, Kuppuswamy M, Venkatesh L, Srinivasan A, Chinnadurai G. Functional substitution of the basic domain of the HIV-1 trans-activator, Tat, with the basic domain of the functionally heterologous Rev. Virology. 1990 May;176(1):178–183. [PubMed]
  • Weeks KM, Ampe C, Schultz SC, Steitz TA, Crothers DM. Fragments of the HIV-1 Tat protein specifically bind TAR RNA. Science. 1990 Sep 14;249(4974):1281–1285. [PubMed]
  • Wright CM, Felber BK, Paskalis H, Pavlakis GN. Expression and characterization of the trans-activator of HTLV-III/LAV virus. Science. 1986 Nov 21;234(4779):988–992. [PubMed]

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