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. 1994 Jul 25;22(14):2740–2743. doi: 10.1093/nar/22.14.2740

Yeast BTF3 protein is encoded by duplicated genes and inhibits the expression of some genes in vivo.

G Z Hu 1, H Ronne 1
PMCID: PMC308242  PMID: 8052529

Abstract

BTF3 is a human protein that is thought to be involved in transcription by RNA polymerase II [Zheng et al., Cell 50, 361-368, 1987]. A yeast homologue of BTF3, Egd1p, has been identified by its ability to enhance DNA binding of the Gal4p activator [Parthun et al., Mol. Cell. Biol. 12, 5683-5689, 1992]. We have cloned a second yeast gene, BTT1, which also encodes a BTF3 homologue. Btt1p and Egd1p are highly similar in sequence, which suggests that they are duplicated proteins with similar functions. Gene disruptions were used to investigate the function of the two proteins. Consistent with published results, we found that loss of EGD1 causes a minor defect in GAL gene induction. Loss of BTT1 has little if any effect. Surprisingly, we found that cells which lack both genes instead express the GAL1 and GAL10 mRNAs at much higher levels than wild type cells. This suggests that BTF3 really plays a negative role in GAL gene expression. Further experiments revealed that expression of the ACT1 and SSO1 genes also is elevated in cells that lack EGD1 and BTT1. In contrast, expression of rRNA and tRNA was not affected. We conclude that Btt1p and Egd1p have redundant functions in vivo, and that they exert a negative effect on the expression of several genes that are transcribed by RNA polymerase II.

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Selected References

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

  1. Aalto M. K., Ronne H., Keränen S. Yeast syntaxins Sso1p and Sso2p belong to a family of related membrane proteins that function in vesicular transport. EMBO J. 1993 Nov;12(11):4095–4104. doi: 10.1002/j.1460-2075.1993.tb06093.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Citron B. A., Donelson J. E. Sequence of the Saccharomyces GAL region and its transcription in vivo. J Bacteriol. 1984 Apr;158(1):269–278. doi: 10.1128/jb.158.1.269-278.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Drapkin R., Merino A., Reinberg D. Regulation of RNA polymerase II transcription. Curr Opin Cell Biol. 1993 Jun;5(3):469–476. doi: 10.1016/0955-0674(93)90013-g. [DOI] [PubMed] [Google Scholar]
  4. Emilsson V., Kurland C. G. Growth rate dependence of transfer RNA abundance in Escherichia coli. EMBO J. 1990 Dec;9(13):4359–4366. doi: 10.1002/j.1460-2075.1990.tb07885.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fascher K. D., Schmitz J., Hörz W. Role of trans-activating proteins in the generation of active chromatin at the PHO5 promoter in S. cerevisiae. EMBO J. 1990 Aug;9(8):2523–2528. doi: 10.1002/j.1460-2075.1990.tb07432.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Flores O., Lu H., Reinberg D. Factors involved in specific transcription by mammalian RNA polymerase II. Identification and characterization of factor IIH. J Biol Chem. 1992 Feb 5;267(4):2786–2793. [PubMed] [Google Scholar]
  7. Gallwitz D., Sures I. Structure of a split yeast gene: complete nucleotide sequence of the actin gene in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1980 May;77(5):2546–2550. doi: 10.1073/pnas.77.5.2546. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gill G. Complexes with a common core. Curr Biol. 1992 Oct;2(10):565–567. doi: 10.1016/0960-9822(92)90046-d. [DOI] [PubMed] [Google Scholar]
  9. Greenblatt J. Roles of TFIID in transcriptional initiation by RNA polymerase II. Cell. 1991 Sep 20;66(6):1067–1070. doi: 10.1016/0092-8674(91)90027-v. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Hansen J. C., Wolffe A. P. A role for histones H2A/H2B in chromatin folding and transcriptional repression. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2339–2343. doi: 10.1073/pnas.91.6.2339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hu G. Z., Ronne H. Overexpression of yeast PAM1 gene permits survival without protein phosphatase 2A and induces a filamentous phenotype. J Biol Chem. 1994 Feb 4;269(5):3429–3435. [PubMed] [Google Scholar]
  13. Johnson A. D., Herskowitz I. A repressor (MAT alpha 2 Product) and its operator control expression of a set of cell type specific genes in yeast. Cell. 1985 Aug;42(1):237–247. doi: 10.1016/s0092-8674(85)80119-7. [DOI] [PubMed] [Google Scholar]
  14. Johnston M., Davis R. W. Sequences that regulate the divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae. Mol Cell Biol. 1984 Aug;4(8):1440–1448. doi: 10.1128/mcb.4.8.1440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nehlin J. O., Carlberg M., Ronne H. Control of yeast GAL genes by MIG1 repressor: a transcriptional cascade in the glucose response. EMBO J. 1991 Nov;10(11):3373–3377. doi: 10.1002/j.1460-2075.1991.tb04901.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nehlin J. O., Carlberg M., Ronne H. Yeast SKO1 gene encodes a bZIP protein that binds to the CRE motif and acts as a repressor of transcription. Nucleic Acids Res. 1992 Oct 25;20(20):5271–5278. doi: 10.1093/nar/20.20.5271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nehlin J. O., Carlberg M., Ronne H. Yeast galactose permease is related to yeast and mammalian glucose transporters. Gene. 1989 Dec 28;85(2):313–319. doi: 10.1016/0378-1119(89)90423-x. [DOI] [PubMed] [Google Scholar]
  18. Nehlin J. O., Ronne H. Yeast MIG1 repressor is related to the mammalian early growth response and Wilms' tumour finger proteins. EMBO J. 1990 Sep;9(9):2891–2898. doi: 10.1002/j.1460-2075.1990.tb07479.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Parthun M. R., Mangus D. A., Jaehning J. A. The EGD1 product, a yeast homolog of human BTF3, may be involved in GAL4 DNA binding. Mol Cell Biol. 1992 Dec;12(12):5683–5689. doi: 10.1128/mcb.12.12.5683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ronne H., Carlberg M., Hu G. Z., Nehlin J. O. Protein phosphatase 2A in Saccharomyces cerevisiae: effects on cell growth and bud morphogenesis. Mol Cell Biol. 1991 Oct;11(10):4876–4884. doi: 10.1128/mcb.11.10.4876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ronne H., Rothstein R. Mitotic sectored colonies: evidence of heteroduplex DNA formation during direct repeat recombination. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2696–2700. doi: 10.1073/pnas.85.8.2696. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Roth S. Y., Shimizu M., Johnson L., Grunstein M., Simpson R. T. Stable nucleosome positioning and complete repression by the yeast alpha 2 repressor are disrupted by amino-terminal mutations in histone H4. Genes Dev. 1992 Mar;6(3):411–425. doi: 10.1101/gad.6.3.411. [DOI] [PubMed] [Google Scholar]
  23. Rothstein R. J. One-step gene disruption in yeast. Methods Enzymol. 1983;101:202–211. doi: 10.1016/0076-6879(83)01015-0. [DOI] [PubMed] [Google Scholar]
  24. Rubtsov P. M., Musakhanov M. M., Zakharyev V. M., Krayev A. S., Skryabin K. G., Bayev A. A. The structure of the yeast ribosomal RNA genes. I. The complete nucleotide sequence of the 18S ribosomal RNA gene from Saccharomyces cerevisiae. Nucleic Acids Res. 1980 Dec 11;8(23):5779–5794. doi: 10.1093/nar/8.23.5779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sawadogo M., Sentenac A. RNA polymerase B (II) and general transcription factors. Annu Rev Biochem. 1990;59:711–754. doi: 10.1146/annurev.bi.59.070190.003431. [DOI] [PubMed] [Google Scholar]
  26. Schlesser A., Ulaszewski S., Ghislain M., Goffeau A. A second transport ATPase gene in Saccharomyces cerevisiae. J Biol Chem. 1988 Dec 25;263(36):19480–19487. [PubMed] [Google Scholar]
  27. Struhl K. Constitutive and inducible Saccharomyces cerevisiae promoters: evidence for two distinct molecular mechanisms. Mol Cell Biol. 1986 Nov;6(11):3847–3853. doi: 10.1128/mcb.6.11.3847. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Thomas B. J., Rothstein R. Elevated recombination rates in transcriptionally active DNA. Cell. 1989 Feb 24;56(4):619–630. doi: 10.1016/0092-8674(89)90584-9. [DOI] [PubMed] [Google Scholar]
  29. Toda T., Cameron S., Sass P., Zoller M., Wigler M. Three different genes in S. cerevisiae encode the catalytic subunits of the cAMP-dependent protein kinase. Cell. 1987 Jul 17;50(2):277–287. doi: 10.1016/0092-8674(87)90223-6. [DOI] [PubMed] [Google Scholar]
  30. Verdier J. M. Regulatory DNA-binding proteins in yeast: an overview. Yeast. 1990 Jul-Aug;6(4):271–297. doi: 10.1002/yea.320060402. [DOI] [PubMed] [Google Scholar]
  31. White R. J., Jackson S. P. The TATA-binding protein: a central role in transcription by RNA polymerases I, II and III. Trends Genet. 1992 Aug;8(8):284–288. doi: 10.1016/0168-9525(92)90255-3. [DOI] [PubMed] [Google Scholar]
  32. Zheng X. M., Black D., Chambon P., Egly J. M. Sequencing and expression of complementary DNA for the general transcription factor BTF3. Nature. 1990 Apr 5;344(6266):556–559. doi: 10.1038/344556a0. [DOI] [PubMed] [Google Scholar]
  33. Zheng X. M., Moncollin V., Egly J. M., Chambon P. A general transcription factor forms a stable complex with RNA polymerase B (II). Cell. 1987 Jul 31;50(3):361–368. doi: 10.1016/0092-8674(87)90490-9. [DOI] [PubMed] [Google Scholar]

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