Abstract
Saccharomyces cerevisiae responds to galactose as the sole source of carbon by activating the GAL genes encoding the enzymes of the Leloir pathway. Here, we show in vitro that the switch from repressed to activated gene expression involves the interplay of three proteins [an activator (Gal4p), a repressor (Gal80p) and an inducer (Gal3p)] and two small molecules (galactose and ATP). We also show that the galactose- and ATP-dependent interaction between Gal3p and Gal80p occurs without disruption of the Gal80p-Gal4p interaction. Thus, Gal3p-mediated activation of transcription occurs via the formation of a tripartite protein complex.
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- Bhat P. J., Hopper J. E. Overproduction of the GAL1 or GAL3 protein causes galactose-independent activation of the GAL4 protein: evidence for a new model of induction for the yeast GAL/MEL regulon. Mol Cell Biol. 1992 Jun;12(6):2701–2707. doi: 10.1128/mcb.12.6.2701-2707.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bhat P. J., Hopper J. E. The mechanism of inducer formation in gal3 mutants of the yeast galactose system is independent of normal galactose metabolism and mitochondrial respiratory function. Genetics. 1991 Jun;128(2):233–239. doi: 10.1093/genetics/128.2.233. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bhat P. J., Oh D., Hopper J. E. Analysis of the GAL3 signal transduction pathway activating GAL4 protein-dependent transcription in Saccharomyces cerevisiae. Genetics. 1990 Jun;125(2):281–291. doi: 10.1093/genetics/125.2.281. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Blank T. E., Woods M. P., Lebo C. M., Xin P., Hopper J. E. Novel Gal3 proteins showing altered Gal80p binding cause constitutive transcription of Gal4p-activated genes in Saccharomyces cerevisiae. Mol Cell Biol. 1997 May;17(5):2566–2575. doi: 10.1128/mcb.17.5.2566. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Giniger E., Varnum S. M., Ptashne M. Specific DNA binding of GAL4, a positive regulatory protein of yeast. Cell. 1985 Apr;40(4):767–774. doi: 10.1016/0092-8674(85)90336-8. [DOI] [PubMed] [Google Scholar]
- Hashimoto H., Kikuchi Y., Nogi Y., Fukasawa T. Regulation of expression of the galactose gene cluster in Saccharomyces cerevisiae. Isolation and characterization of the regulatory gene GAL4. Mol Gen Genet. 1983;191(1):31–38. doi: 10.1007/BF00330886. [DOI] [PubMed] [Google Scholar]
- 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]
- Leuther K. K., Johnston S. A. Nondissociation of GAL4 and GAL80 in vivo after galactose induction. Science. 1992 May 29;256(5061):1333–1335. doi: 10.1126/science.1598579. [DOI] [PubMed] [Google Scholar]
- Lillie J. W., Green M., Green M. R. An adenovirus E1a protein region required for transformation and transcriptional repression. Cell. 1986 Sep 26;46(7):1043–1051. doi: 10.1016/0092-8674(86)90704-x. [DOI] [PubMed] [Google Scholar]
- 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]
- Lohr D., Venkov P., Zlatanova J. Transcriptional regulation in the yeast GAL gene family: a complex genetic network. FASEB J. 1995 Jun;9(9):777–787. doi: 10.1096/fasebj.9.9.7601342. [DOI] [PubMed] [Google Scholar]
- Lue N. F., Chasman D. I., Buchman A. R., Kornberg R. D. Interaction of GAL4 and GAL80 gene regulatory proteins in vitro. Mol Cell Biol. 1987 Oct;7(10):3446–3451. doi: 10.1128/mcb.7.10.3446-3451.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nogi Y., Fukasawa T. Nucleotide sequence of the yeast regulatory gene GAL80. Nucleic Acids Res. 1984 Dec 21;12(24):9287–9298. doi: 10.1093/nar/12.24.9287. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nogi Y., Shimada H., Matsuzaki Y., Hashimoto H., Fukasawa T. Regulation of expression of the galactose gene cluster in Saccharomyces cerevisiae. II. The isolation and dosage effect of the regulatory gene GAL80. Mol Gen Genet. 1984;195(1-2):29–34. doi: 10.1007/BF00332719. [DOI] [PubMed] [Google Scholar]
- Ohashi Y., Brickman J. M., Furman E., Middleton B., Carey M. Modulating the potency of an activator in a yeast in vitro transcription system. Mol Cell Biol. 1994 Apr;14(4):2731–2739. doi: 10.1128/mcb.14.4.2731-2739.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- ROTMAN B., SPIEGELMAN S. The conversion of negatives to positives in slow adapting populations of yeast. J Bacteriol. 1953 Oct;66(4):492–497. doi: 10.1128/jb.66.4.492-497.1953. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reece R. J., Platt A. Signaling activation and repression of RNA polymerase II transcription in yeast. Bioessays. 1997 Nov;19(11):1001–1010. doi: 10.1002/bies.950191110. [DOI] [PubMed] [Google Scholar]
- Reece R. J., Ptashne M. Determinants of binding-site specificity among yeast C6 zinc cluster proteins. Science. 1993 Aug 13;261(5123):909–911. doi: 10.1126/science.8346441. [DOI] [PubMed] [Google Scholar]
- Reece R. J., Rickles R. J., Ptashne M. Overproduction and single-step purification of GAL4 fusion proteins from Escherichia coli. Gene. 1993 Apr 15;126(1):105–107. doi: 10.1016/0378-1119(93)90596-u. [DOI] [PubMed] [Google Scholar]
- Suzuki-Fujimoto T., Fukuma M., Yano K. I., Sakurai H., Vonika A., Johnston S. A., Fukasawa T. Analysis of the galactose signal transduction pathway in Saccharomyces cerevisiae: interaction between Gal3p and Gal80p. Mol Cell Biol. 1996 May;16(5):2504–2508. doi: 10.1128/mcb.16.5.2504. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wu Y., Reece R. J., Ptashne M. Quantitation of putative activator-target affinities predicts transcriptional activating potentials. EMBO J. 1996 Aug 1;15(15):3951–3963. [PMC free article] [PubMed] [Google Scholar]
- Yano K., Fukasawa T. Galactose-dependent reversible interaction of Gal3p with Gal80p in the induction pathway of Gal4p-activated genes of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1997 Mar 4;94(5):1721–1726. doi: 10.1073/pnas.94.5.1721. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zenke F. T., Engles R., Vollenbroich V., Meyer J., Hollenberg C. P., Breunig K. D. Activation of Gal4p by galactose-dependent interaction of galactokinase and Gal80p. Science. 1996 Jun 14;272(5268):1662–1665. doi: 10.1126/science.272.5268.1662. [DOI] [PubMed] [Google Scholar]
