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. 1995 Aug 15;92(17):7677–7680. doi: 10.1073/pnas.92.17.7677

A single GAL4 dimer can maximally activate transcription under physiological conditions.

H E Xu 1, T Kodadek 1, S A Johnston 1
PMCID: PMC41208  PMID: 7644476

Abstract

Most eukaryotic promoters contain multiple binding sites for one or more transcriptional activators that interact in a synergistic manner. A common view is that synergism is a manifestation of the need for many contacts between activators and the general transcription machinery that a single activator presumably cannot fulfill. In this model, various combinations of protein-protein interactions control the level of gene expression. However, we show here that under physiological conditions, a single binding site and presumably GAL4 can activate transcription to the maximum possible level in vivo. Synergistic effects in this natural system are shown to be consistent with cooperative DNA binding. These results point to DNA occupancy as the major element in fine tuning gene expression in the galactose regulon.

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Selected References

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

  1. Allan G. F., Ing N. H., Tsai S. Y., Srinivasan G., Weigel N. L., Thompson E. B., Tsai M. J., O'Malley B. W. Synergism between steroid response and promoter elements during cell-free transcription. J Biol Chem. 1991 Mar 25;266(9):5905–5910. [PubMed] [Google Scholar]
  2. Baniahmad C., Muller M., Altschmied J., Renkawitz R. Co-operative binding of the glucocorticoid receptor DNA binding domain is one of at least two mechanisms for synergism. J Mol Biol. 1991 Nov 20;222(2):155–165. doi: 10.1016/0022-2836(91)90202-h. [DOI] [PubMed] [Google Scholar]
  3. Carey M., Kakidani H., Leatherwood J., Mostashari F., Ptashne M. An amino-terminal fragment of GAL4 binds DNA as a dimer. J Mol Biol. 1989 Oct 5;209(3):423–432. doi: 10.1016/0022-2836(89)90007-7. [DOI] [PubMed] [Google Scholar]
  4. Carey M., Kolman J., Katz D. A., Gradoville L., Barberis L., Miller G. Transcriptional synergy by the Epstein-Barr virus transactivator ZEBRA. J Virol. 1992 Aug;66(8):4803–4813. doi: 10.1128/jvi.66.8.4803-4813.1992. [DOI] [PMC free article] [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. 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]
  7. Choy B., Green M. R. Eukaryotic activators function during multiple steps of preinitiation complex assembly. Nature. 1993 Dec 9;366(6455):531–536. doi: 10.1038/366531a0. [DOI] [PubMed] [Google Scholar]
  8. Courey A. J., Holtzman D. A., Jackson S. P., Tjian R. Synergistic activation by the glutamine-rich domains of human transcription factor Sp1. Cell. 1989 Dec 1;59(5):827–836. doi: 10.1016/0092-8674(89)90606-5. [DOI] [PubMed] [Google Scholar]
  9. Du W., Thanos D., Maniatis T. Mechanisms of transcriptional synergism between distinct virus-inducible enhancer elements. Cell. 1993 Sep 10;74(5):887–898. doi: 10.1016/0092-8674(93)90468-6. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Giniger E., Ptashne M. Cooperative DNA binding of the yeast transcriptional activator GAL4. Proc Natl Acad Sci U S A. 1988 Jan;85(2):382–386. doi: 10.1073/pnas.85.2.382. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Griggs D. W., Johnston M. Regulated expression of the GAL4 activator gene in yeast provides a sensitive genetic switch for glucose repression. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8597–8601. doi: 10.1073/pnas.88.19.8597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Herschlag D., Johnson F. B. Synergism in transcriptional activation: a kinetic view. Genes Dev. 1993 Feb;7(2):173–179. doi: 10.1101/gad.7.2.173. [DOI] [PubMed] [Google Scholar]
  14. Johnston M. A model fungal gene regulatory mechanism: the GAL genes of Saccharomyces cerevisiae. Microbiol Rev. 1987 Dec;51(4):458–476. doi: 10.1128/mr.51.4.458-476.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Johnston S. A., Hopper J. E. Isolation of the yeast regulatory gene GAL4 and analysis of its dosage effects on the galactose/melibiose regulon. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6971–6975. doi: 10.1073/pnas.79.22.6971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Kang T., Martins T., Sadowski I. Wild type GAL4 binds cooperatively to the GAL1-10 UASG in vitro. J Biol Chem. 1993 May 5;268(13):9629–9635. [PubMed] [Google Scholar]
  18. Leuther K. K., Salmeron J. M., Johnston S. A. Genetic evidence that an activation domain of GAL4 does not require acidity and may form a beta sheet. Cell. 1993 Feb 26;72(4):575–585. doi: 10.1016/0092-8674(93)90076-3. [DOI] [PubMed] [Google Scholar]
  19. Li R., Knight J. D., Jackson S. P., Tjian R., Botchan M. R. Direct interaction between Sp1 and the BPV enhancer E2 protein mediates synergistic activation of transcription. Cell. 1991 May 3;65(3):493–505. doi: 10.1016/0092-8674(91)90467-d. [DOI] [PubMed] [Google Scholar]
  20. Lin Y. S., Carey M., Ptashne M., Green M. R. How different eukaryotic transcriptional activators can cooperate promiscuously. Nature. 1990 May 24;345(6273):359–361. doi: 10.1038/345359a0. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Muller M., Baniahmad C., Kaltschmidt C., Renkawitz R. Multiple domains of the glucocorticoid receptor involved in synergism with the CACCC box factor(s). Mol Endocrinol. 1991 Oct;5(10):1498–1503. doi: 10.1210/mend-5-10-1498. [DOI] [PubMed] [Google Scholar]
  23. Oliviero S., Struhl K. Synergistic transcriptional enhancement does not depend on the number of acidic activation domains bound to the promoter. Proc Natl Acad Sci U S A. 1991 Jan 1;88(1):224–228. doi: 10.1073/pnas.88.1.224. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pascal E., Tjian R. Different activation domains of Sp1 govern formation of multimers and mediate transcriptional synergism. Genes Dev. 1991 Sep;5(9):1646–1656. doi: 10.1101/gad.5.9.1646. [DOI] [PubMed] [Google Scholar]
  25. Sadowski I., Ma J., Triezenberg S., Ptashne M. GAL4-VP16 is an unusually potent transcriptional activator. Nature. 1988 Oct 6;335(6190):563–564. doi: 10.1038/335563a0. [DOI] [PubMed] [Google Scholar]
  26. Schatt M. D., Rusconi S., Schaffner W. A single DNA-binding transcription factor is sufficient for activation from a distant enhancer and/or from a promoter position. EMBO J. 1990 Feb;9(2):481–487. doi: 10.1002/j.1460-2075.1990.tb08134.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schüle R., Muller M., Kaltschmidt C., Renkawitz R. Many transcription factors interact synergistically with steroid receptors. Science. 1988 Dec 9;242(4884):1418–1420. doi: 10.1126/science.3201230. [DOI] [PubMed] [Google Scholar]
  28. Svaren J., Wineinger B. D., Chalkley R. Extent of in vivo binding by an upstream activation factor and the role of multiple binding sites in synergistic transcriptional activation. J Biol Chem. 1994 Aug 12;269(32):20771–20779. [PubMed] [Google Scholar]
  29. Taylor I. C., Workman J. L., Schuetz T. J., Kingston R. E. Facilitated binding of GAL4 and heat shock factor to nucleosomal templates: differential function of DNA-binding domains. Genes Dev. 1991 Jul;5(7):1285–1298. doi: 10.1101/gad.5.7.1285. [DOI] [PubMed] [Google Scholar]
  30. Tsai S. Y., Tsai M. J., O'Malley B. W. Cooperative binding of steroid hormone receptors contributes to transcriptional synergism at target enhancer elements. Cell. 1989 May 5;57(3):443–448. doi: 10.1016/0092-8674(89)90919-7. [DOI] [PubMed] [Google Scholar]
  31. Vashee S., Xu H., Johnston S. A., Kodadek T. How do "Zn2 cys6" proteins distinguish between similar upstream activation sites? Comparison of the DNA-binding specificity of the GAL4 protein in vitro and in vivo. J Biol Chem. 1993 Nov 25;268(33):24699–24706. [PubMed] [Google Scholar]
  32. Wang W. D., Gralla J. D. Differential ability of proximal and remote element pairs to cooperate in activating RNA polymerase II transcription. Mol Cell Biol. 1991 Sep;11(9):4561–4571. doi: 10.1128/mcb.11.9.4561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. White J., Brou C., Wu J., Lutz Y., Moncollin V., Chambon P. The acidic transcriptional activator GAL-VP16 acts on preformed template-committed complexes. EMBO J. 1992 Jun;11(6):2229–2240. doi: 10.1002/j.1460-2075.1992.tb05282.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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