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. 1993 Feb 15;90(4):1571–1575. doi: 10.1073/pnas.90.4.1571

Electrostatic interactions modulate the RNA-binding and transactivation specificities of the human immunodeficiency virus and simian immunodeficiency virus Tat proteins.

J Tao 1, A D Frankel 1
PMCID: PMC45916  PMID: 8434019

Abstract

The transcriptional activating (Tat) proteins from human immunodeficiency virus and simian immunodeficiency virus are sequence-specific RNA-binding proteins. In human immunodeficiency virus Tat, a single arginine residue, flanked on each side by three to four basic amino acids, mediates specific binding to a bulge region in trans-acting responsive element (TAR) RNA. We have systematically mutated the flanking charged residues and found that, in addition to the position of the sequence-specific arginine, the particular arrangement of nonspecific electrostatic interactions is an important determinant of RNA-binding specificity and transactivation activity. These additional electrostatic contacts may help stabilize the structure of TAR RNA when bound to arginine. One critical electrostatic interaction, located two residues N-terminal to the arginine, is absent in the simian immunodeficiency virus Tat protein and accounts for the difference in promoter specificities of the human and simian immunodeficiency viral proteins.

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Selected References

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

  1. Arya S. K., Gallo R. C. Human immunodeficiency virus type 2 long terminal repeat: analysis of regulatory elements. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9753–9757. doi: 10.1073/pnas.85.24.9753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berkhout B., Gatignol A., Silver J., Jeang K. T. Efficient trans-activation by the HIV-2 Tat protein requires a duplicated TAR RNA structure. Nucleic Acids Res. 1990 Apr 11;18(7):1839–1846. doi: 10.1093/nar/18.7.1839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Calnan B. J., Biancalana S., Hudson D., Frankel A. D. Analysis of arginine-rich peptides from the HIV Tat protein reveals unusual features of RNA-protein recognition. Genes Dev. 1991 Feb;5(2):201–210. doi: 10.1101/gad.5.2.201. [DOI] [PubMed] [Google Scholar]
  4. Carroll R., Peterlin B. M., Derse D. Inhibition of human immunodeficiency virus type 1 Tat activity by coexpression of heterologous trans activators. J Virol. 1992 Apr;66(4):2000–2007. doi: 10.1128/jvi.66.4.2000-2007.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Cordingley M. G., LaFemina R. L., Callahan P. L., Condra J. H., Sardana V. V., Graham D. J., Nguyen T. M., LeGrow K., Gotlib L., Schlabach A. J. 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. doi: 10.1073/pnas.87.22.8985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dang C. V., Lee W. M. Nuclear and nucleolar targeting sequences of c-erb-A, c-myb, N-myc, p53, HSP70, and HIV tat proteins. J Biol Chem. 1989 Oct 25;264(30):18019–18023. [PubMed] [Google Scholar]
  8. Delling U., Roy S., Sumner-Smith M., Barnett R., Reid L., Rosen C. A., Sonenberg N. The number of positively charged amino acids in the basic domain of Tat is critical for trans-activation and complex formation with TAR RNA. Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6234–6238. doi: 10.1073/pnas.88.14.6234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dingwall C., Ernberg I., Gait M. J., Green S. M., Heaphy S., Karn J., Lowe A. D., Singh M., Skinner M. A. HIV-1 tat protein stimulates transcription by binding to a U-rich bulge in the stem of the TAR RNA structure. EMBO J. 1990 Dec;9(12):4145–4153. doi: 10.1002/j.1460-2075.1990.tb07637.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Elangovan B., Subramanian T., Chinnadurai G. Functional comparison of the basic domains of the Tat proteins of human immunodeficiency virus types 1 and 2 in trans activation. J Virol. 1992 Apr;66(4):2031–2036. doi: 10.1128/jvi.66.4.2031-2036.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Emerman M., Guyader M., Montagnier L., Baltimore D., Muesing M. A. The specificity of the human immunodeficiency virus type 2 transactivator is different from that of human immunodeficiency virus type 1. EMBO J. 1987 Dec 1;6(12):3755–3760. doi: 10.1002/j.1460-2075.1987.tb02710.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fenrick R., Malim M. H., Hauber J., Le S. Y., Maizel J., Cullen B. R. Functional analysis of the Tat trans activator of human immunodeficiency virus type 2. J Virol. 1989 Dec;63(12):5006–5012. doi: 10.1128/jvi.63.12.5006-5012.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Frankel A. D. Activation of HIV transcription by Tat. Curr Opin Genet Dev. 1992 Apr;2(2):293–298. doi: 10.1016/s0959-437x(05)80287-4. [DOI] [PubMed] [Google Scholar]
  14. Frankel A. D., Pabo C. O. Cellular uptake of the tat protein from human immunodeficiency virus. Cell. 1988 Dec 23;55(6):1189–1193. doi: 10.1016/0092-8674(88)90263-2. [DOI] [PubMed] [Google Scholar]
  15. Hauber J., Malim M. H., Cullen B. R. Mutational analysis of the conserved basic domain of human immunodeficiency virus tat protein. J Virol. 1989 Mar;63(3):1181–1187. doi: 10.1128/jvi.63.3.1181-1187.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lazinski D., Grzadzielska E., Das A. Sequence-specific recognition of RNA hairpins by bacteriophage antiterminators requires a conserved arginine-rich motif. Cell. 1989 Oct 6;59(1):207–218. doi: 10.1016/0092-8674(89)90882-9. [DOI] [PubMed] [Google Scholar]
  17. Puglisi J. D., Tan R., Calnan B. J., Frankel A. D., Williamson J. R. Conformation of the TAR RNA-arginine complex by NMR spectroscopy. Science. 1992 Jul 3;257(5066):76–80. doi: 10.1126/science.1621097. [DOI] [PubMed] [Google Scholar]
  18. Renjifo B., Speck N. A., Winandy S., Hopkins N., Li Y. cis-acting elements in the U3 region of a simian immunodeficiency virus. J Virol. 1990 Jun;64(6):3130–3134. doi: 10.1128/jvi.64.6.3130-3134.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Roy S., Delling U., Chen C. H., Rosen C. A., Sonenberg N. A bulge structure in HIV-1 TAR RNA is required for Tat binding and Tat-mediated trans-activation. Genes Dev. 1990 Aug;4(8):1365–1373. doi: 10.1101/gad.4.8.1365. [DOI] [PubMed] [Google Scholar]
  20. Ruben S., Perkins A., Purcell R., Joung K., Sia R., Burghoff R., Haseltine W. A., Rosen C. A. Structural and functional characterization of human immunodeficiency virus tat protein. J Virol. 1989 Jan;63(1):1–8. doi: 10.1128/jvi.63.1.1-8.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sodroski J., Rosen C., Wong-Staal F., Salahuddin S. Z., Popovic M., Arya S., Gallo R. C., Haseltine W. A. Trans-acting transcriptional regulation of human T-cell leukemia virus type III long terminal repeat. Science. 1985 Jan 11;227(4683):171–173. doi: 10.1126/science.2981427. [DOI] [PubMed] [Google Scholar]
  22. Subramanian T., Govindarajan R., Chinnadurai G. Heterologous basic domain substitutions in the HIV-1 Tat protein reveal an arginine-rich motif required for transactivation. EMBO J. 1991 Aug;10(8):2311–2318. doi: 10.1002/j.1460-2075.1991.tb07768.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tan R., Frankel A. D. Circular dichroism studies suggest that TAR RNA changes conformation upon specific binding of arginine or guanidine. Biochemistry. 1992 Oct 27;31(42):10288–10294. doi: 10.1021/bi00157a016. [DOI] [PubMed] [Google Scholar]
  24. Tao J., Frankel A. D. Specific binding of arginine to TAR RNA. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2723–2726. doi: 10.1073/pnas.89.7.2723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Viglianti G. A., Mullins J. I. Functional comparison of transactivation by simian immunodeficiency virus from rhesus macaques and human immunodeficiency virus type 1. J Virol. 1988 Dec;62(12):4523–4532. doi: 10.1128/jvi.62.12.4523-4532.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Weeks K. M., Ampe C., Schultz S. C., Steitz T. A., Crothers D. M. Fragments of the HIV-1 Tat protein specifically bind TAR RNA. Science. 1990 Sep 14;249(4974):1281–1285. doi: 10.1126/science.2205002. [DOI] [PubMed] [Google Scholar]
  27. Weeks K. M., Crothers D. M. RNA recognition by Tat-derived peptides: interaction in the major groove? Cell. 1991 Aug 9;66(3):577–588. doi: 10.1016/0092-8674(81)90020-9. [DOI] [PubMed] [Google Scholar]

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