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. 1996 Feb 15;24(4):549–557. doi: 10.1093/nar/24.4.549

Activation domains of transcription factors mediate replication dependent transcription from a minimal HIV-1 promoter.

R D Williams 1, B A Lee 1, S P Jackson 1, N J Proudfoot 1
PMCID: PMC145701  PMID: 8604293

Abstract

Transcription from a minimal HIV-1 promoter containing the three Sp1 binding sites and TATA box can be activated without Tat by template DNA replication. Here we show that this activation can also be mediated by recombinant GAL4 fusion proteins containing the activation domains of Sp1, VP16 or CTF (or by full-length GAL4) targeted to the HIV-1 promoter by replacing the Sp1 sites with five GAL4 binding sites. Thus Sp1 is not unique in its ability to mediate replication activated transcription, although the degree of processivity elicited by the different activators varied significantly from strongly processive (GAL4-VP16) to relatively non-processive (GAL4-Sp1 or -CTF). Processive GAL4-VP16-activated transcription, but not efficient initiation, required multiple GAL4 binding sites. In the presence of Tat, transcription with GAL4-SP1 and GAL4-CTF was further activated (principally at the level of processivity) but GAL4-VP16-potentiated transcription was only slightly stimulated. The Tat-dependent switch from non-processive to fully processive transcription was particularly marked for GAL4-Sp1, an effect which may be relevant to the selection of Sp1 binding sites by the HIV-1 promoter.

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Selected References

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

  1. Adams C. C., Workman J. L. Nucleosome displacement in transcription. Cell. 1993 Feb 12;72(3):305–308. doi: 10.1016/0092-8674(93)90109-4. [DOI] [PubMed] [Google Scholar]
  2. Adams S. E., Johnson I. D., Braddock M., Kingsman A. J., Kingsman S. M., Edwards R. M. Synthesis of a gene for the HIV transactivator protein TAT by a novel single stranded approach involving in vivo gap repair. Nucleic Acids Res. 1988 May 25;16(10):4287–4298. doi: 10.1093/nar/16.10.4287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berkhout B., Jeang K. T. Functional roles for the TATA promoter and enhancers in basal and Tat-induced expression of the human immunodeficiency virus type 1 long terminal repeat. J Virol. 1992 Jan;66(1):139–149. doi: 10.1128/jvi.66.1.139-149.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Borowiec J. A., Dean F. B., Bullock P. A., Hurwitz J. Binding and unwinding--how T antigen engages the SV40 origin of DNA replication. Cell. 1990 Jan 26;60(2):181–184. doi: 10.1016/0092-8674(90)90730-3. [DOI] [PubMed] [Google Scholar]
  5. Carey M., Lin Y. S., Green M. R., Ptashne M. A mechanism for synergistic activation of a mammalian gene by GAL4 derivatives. Nature. 1990 May 24;345(6273):361–364. doi: 10.1038/345361a0. [DOI] [PubMed] [Google Scholar]
  6. Cereghini S., Yaniv M. Assembly of transfected DNA into chromatin: structural changes in the origin-promoter-enhancer region upon replication. EMBO J. 1984 Jun;3(6):1243–1253. doi: 10.1002/j.1460-2075.1984.tb01959.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Challberg M. D., Kelly T. J. Animal virus DNA replication. Annu Rev Biochem. 1989;58:671–717. doi: 10.1146/annurev.bi.58.070189.003323. [DOI] [PubMed] [Google Scholar]
  8. Chang C., Gralla J. D. A critical role for chromatin in mounting a synergistic transcriptional response to GAL4-VP16. Mol Cell Biol. 1994 Aug;14(8):5175–5181. doi: 10.1128/mcb.14.8.5175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chang C., Gralla J. D. Properties of initiator-associated transcription mediated by GAL4-VP16. Mol Cell Biol. 1993 Dec;13(12):7469–7475. doi: 10.1128/mcb.13.12.7469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Courey A. J., Tjian R. Analysis of Sp1 in vivo reveals multiple transcriptional domains, including a novel glutamine-rich activation motif. Cell. 1988 Dec 2;55(5):887–898. doi: 10.1016/0092-8674(88)90144-4. [DOI] [PubMed] [Google Scholar]
  11. Emami K. H., Carey M. A synergistic increase in potency of a multimerized VP16 transcriptional activation domain. EMBO J. 1992 Dec;11(13):5005–5012. doi: 10.1002/j.1460-2075.1992.tb05607.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Enver T., Brewer A. C., Patient R. K. Role for DNA replication in beta-globin gene activation. Mol Cell Biol. 1988 Mar;8(3):1301–1308. doi: 10.1128/mcb.8.3.1301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Grass D. S., Read D., Lewis E. D., Manley J. L. Cell- and promoter-specific activation of transcription by DNA replication. Genes Dev. 1987 Dec;1(10):1065–1074. doi: 10.1101/gad.1.10.1065. [DOI] [PubMed] [Google Scholar]
  14. Grosveld G. C., de Boer E., Shewmaker C. K., Flavell R. A. DNA sequences necessary for transcription of the rabbit beta-globin gene in vivo. Nature. 1982 Jan 14;295(5845):120–126. doi: 10.1038/295120a0. [DOI] [PubMed] [Google Scholar]
  15. Jeang K. T., Berkhout B., Dropulic B. Effects of integration and replication on transcription of the HIV-1 long terminal repeat. J Biol Chem. 1993 Nov 25;268(33):24940–24949. [PubMed] [Google Scholar]
  16. Jeang K. T., Chun R., Lin N. H., Gatignol A., Glabe C. G., Fan H. In vitro and in vivo binding of human immunodeficiency virus type 1 Tat protein and Sp1 transcription factor. J Virol. 1993 Oct;67(10):6224–6233. doi: 10.1128/jvi.67.10.6224-6233.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jones K. A., Peterlin B. M. Control of RNA initiation and elongation at the HIV-1 promoter. Annu Rev Biochem. 1994;63:717–743. doi: 10.1146/annurev.bi.63.070194.003441. [DOI] [PubMed] [Google Scholar]
  18. Kamakaka R. T., Bulger M., Kadonaga J. T. Potentiation of RNA polymerase II transcription by Gal4-VP16 during but not after DNA replication and chromatin assembly. Genes Dev. 1993 Sep;7(9):1779–1795. doi: 10.1101/gad.7.9.1779. [DOI] [PubMed] [Google Scholar]
  19. 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. doi: 10.1128/jvi.66.6.3932-3936.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kessler M., Mathews M. B. Tat transactivation of the human immunodeficiency virus type 1 promoter is influenced by basal promoter activity and the simian virus 40 origin of DNA replication. Proc Natl Acad Sci U S A. 1991 Nov 15;88(22):10018–10022. doi: 10.1073/pnas.88.22.10018. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Krumm A., Hickey L. B., Groudine M. Promoter-proximal pausing of RNA polymerase II defines a general rate-limiting step after transcription initiation. Genes Dev. 1995 Mar 1;9(5):559–572. doi: 10.1101/gad.9.5.559. [DOI] [PubMed] [Google Scholar]
  22. Lewis P., Hensel M., Emerman M. Human immunodeficiency virus infection of cells arrested in the cell cycle. EMBO J. 1992 Aug;11(8):3053–3058. doi: 10.1002/j.1460-2075.1992.tb05376.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lin Y. S., Carey M. F., Ptashne M., Green M. R. GAL4 derivatives function alone and synergistically with mammalian activators in vitro. Cell. 1988 Aug 26;54(5):659–664. doi: 10.1016/s0092-8674(88)80010-2. [DOI] [PubMed] [Google Scholar]
  24. Ma J., Ptashne M. Deletion analysis of GAL4 defines two transcriptional activating segments. Cell. 1987 Mar 13;48(5):847–853. doi: 10.1016/0092-8674(87)90081-x. [DOI] [PubMed] [Google Scholar]
  25. Madore S. J., Cullen B. R. Functional similarities between HIV-1 Tat and DNA sequence-specific transcriptional activators. Virology. 1995 Feb 1;206(2):1150–1154. doi: 10.1006/viro.1995.1041. [DOI] [PubMed] [Google Scholar]
  26. Martinez E., Dusserre Y., Wahli W., Mermod N. Synergistic transcriptional activation by CTF/NF-I and the estrogen receptor involves stabilized interactions with a limiting target factor. Mol Cell Biol. 1991 Jun;11(6):2937–2945. doi: 10.1128/mcb.11.6.2937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Matsui T., Murayama M., Mita T. Adenovirus 2 peptide IX gene is expressed only on replicated DNA molecules. Mol Cell Biol. 1986 Dec;6(12):4149–4154. doi: 10.1128/mcb.6.12.4149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. 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. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Murphy S., Yoon J. B., Gerster T., Roeder R. G. Oct-1 and Oct-2 potentiate functional interactions of a transcription factor with the proximal sequence element of small nuclear RNA genes. Mol Cell Biol. 1992 Jul;12(7):3247–3261. doi: 10.1128/mcb.12.7.3247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nahreini P., Mathews M. B. Effects of the simian virus 40 origin of replication on transcription from the human immunodeficiency virus type 1 promoter. J Virol. 1995 Feb;69(2):1296–1301. doi: 10.1128/jvi.69.2.1296-1301.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Proudfoot N. J., Lee B. A., Monks J. Multiple SP1 binding sites confer enhancer-independent, replication-activated transcription of HIV-1 and globin gene promoters. New Biol. 1992 Apr;4(4):369–381. [PubMed] [Google Scholar]
  32. Sadowski I., Ptashne M. A vector for expressing GAL4(1-147) fusions in mammalian cells. Nucleic Acids Res. 1989 Sep 25;17(18):7539–7539. doi: 10.1093/nar/17.18.7539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schuitemaker H., Kootstra N. A., Fouchier R. A., Hooibrink B., Miedema F. Productive HIV-1 infection of macrophages restricted to the cell fraction with proliferative capacity. EMBO J. 1994 Dec 15;13(24):5929–5936. doi: 10.1002/j.1460-2075.1994.tb06938.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Seipel K., Georgiev O., Schaffner W. Different activation domains stimulate transcription from remote ('enhancer') and proximal ('promoter') positions. EMBO J. 1992 Dec;11(13):4961–4968. doi: 10.1002/j.1460-2075.1992.tb05603.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Southgate C. D., Green M. R. The HIV-1 Tat protein activates transcription from an upstream DNA-binding site: implications for Tat function. Genes Dev. 1991 Dec;5(12B):2496–2507. doi: 10.1101/gad.5.12b.2496. [DOI] [PubMed] [Google Scholar]
  36. Thomas G. P., Mathews M. B. DNA replication and the early to late transition in adenovirus infection. Cell. 1980 Nov;22(2 Pt 2):523–533. doi: 10.1016/0092-8674(80)90362-1. [DOI] [PubMed] [Google Scholar]
  37. Treisman R., Green M. R., Maniatis T. cis and trans activation of globin gene transcription in transient assays. Proc Natl Acad Sci U S A. 1983 Dec;80(24):7428–7432. doi: 10.1073/pnas.80.24.7428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Venkatesh L. K., Chinnadurai G. Activation of the adenovirus 2 protein IX promoter by DNA replication in a transient expression assay. Nucleic Acids Res. 1987 Mar 11;15(5):2235–2250. doi: 10.1093/nar/15.5.2235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Webster N., Jin J. R., Green S., Hollis M., Chambon P. The yeast UASG is a transcriptional enhancer in human HeLa cells in the presence of the GAL4 trans-activator. Cell. 1988 Jan 29;52(2):169–178. doi: 10.1016/0092-8674(88)90505-3. [DOI] [PubMed] [Google Scholar]
  40. Whitelaw E., Hogben P., Hanscombe O., Proudfoot N. J. Transcriptional promiscuity of the human alpha-globin gene. Mol Cell Biol. 1989 Jan;9(1):241–251. doi: 10.1128/mcb.9.1.241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wilson A. C., Patient R. K. DNA replication facilitates the action of transcriptional enhancers in transient expression assays. Nucleic Acids Res. 1993 Sep 11;21(18):4296–4304. doi: 10.1093/nar/21.18.4296. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wolffe A. P. Implications of DNA replication for eukaryotic gene expression. J Cell Sci. 1991 Jun;99(Pt 2):201–206. doi: 10.1242/jcs.99.2.201. [DOI] [PubMed] [Google Scholar]
  43. Yankulov K., Blau J., Purton T., Roberts S., Bentley D. L. Transcriptional elongation by RNA polymerase II is stimulated by transactivators. Cell. 1994 Jun 3;77(5):749–759. doi: 10.1016/0092-8674(94)90058-2. [DOI] [PubMed] [Google Scholar]

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