Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1993 Apr;13(4):2081–2090. doi: 10.1128/mcb.13.4.2081

Three downstream sites repress transcription of a Ty2 retrotransposon in Saccharomyces cerevisiae.

P J Farabaugh 1, A Vimaladithan 1, S Türkel 1, R Johnson 1, H Zhao 1
PMCID: PMC359529  PMID: 8384303

Abstract

Transcription of Ty1 and Ty2 retrotransposons of the yeast Saccharomyces cerevisiae is modulated by multiple downstream regulatory sites. Both transposon families include a positively acting site within the transcribed region which resembles a higher eukaryotic enhancer. We have demonstrated the existence of a repression site distal to the enhancer of the Ty2-917 element. Here we describe experiments investigating the internal structure of this site. We show that this 200-bp region includes three distinct repression sites which we term DRSI (downstream repression site I), DRSII, and DRSIII. Individually each site causes almost twofold repression, and together the sites repress eightfold. Unexpectedly, when the entire region encompassing the DRS sites is moved outside the transcription unit, it acts as a qualitatively positively acting element. In this context the DRS sites still repress transcription, since eliminating them increases transcription further. That the region can activate transcription implies that it includes activation sites in addition to the three repression sites. The change from qualitatively negatively acting to positively acting must reflect a change in the relative effects of the multiple positive and negative sites; when moved outside the transcription unit, the activators predominate. Importantly, DRSII and DRSIII repress transcription autonomously when inserted upstream of a heterologous promoter activated by the transcriptional activator GCN4, showing that they are indeed transcriptional repression sites.

Full text

PDF
2081

Selected References

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

  1. Arndt K. T., Styles C., Fink G. R. Multiple global regulators control HIS4 transcription in yeast. Science. 1987 Aug 21;237(4817):874–880. doi: 10.1126/science.3303332. [DOI] [PubMed] [Google Scholar]
  2. Belcourt M. F., Farabaugh P. J. Ribosomal frameshifting in the yeast retrotransposon Ty: tRNAs induce slippage on a 7 nucleotide minimal site. Cell. 1990 Jul 27;62(2):339–352. doi: 10.1016/0092-8674(90)90371-K. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brand A. H., Breeden L., Abraham J., Sternglanz R., Nasmyth K. Characterization of a "silencer" in yeast: a DNA sequence with properties opposite to those of a transcriptional enhancer. Cell. 1985 May;41(1):41–48. doi: 10.1016/0092-8674(85)90059-5. [DOI] [PubMed] [Google Scholar]
  4. Clare J. J., Belcourt M., Farabaugh P. J. Efficient translational frameshifting occurs within a conserved sequence of the overlap between the two genes of a yeast Ty1 transposon. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6816–6820. doi: 10.1073/pnas.85.18.6816. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clark-Adams C. D., Norris D., Osley M. A., Fassler J. S., Winston F. Changes in histone gene dosage alter transcription in yeast. Genes Dev. 1988 Feb;2(2):150–159. doi: 10.1101/gad.2.2.150. [DOI] [PubMed] [Google Scholar]
  6. Company M., Adler C., Errede B. Identification of a Ty1 regulatory sequence responsive to STE7 and STE12. Mol Cell Biol. 1988 Jun;8(6):2545–2554. doi: 10.1128/mcb.8.6.2545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Company M., Errede B. Cell-type-dependent gene activation by yeast transposon Ty1 involves multiple regulatory determinants. Mol Cell Biol. 1987 Sep;7(9):3205–3211. doi: 10.1128/mcb.7.9.3205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Devlin C., Tice-Baldwin K., Shore D., Arndt K. T. RAP1 is required for BAS1/BAS2- and GCN4-dependent transcription of the yeast HIS4 gene. Mol Cell Biol. 1991 Jul;11(7):3642–3651. doi: 10.1128/mcb.11.7.3642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Durrin L. K., Mann R. K., Grunstein M. Nucleosome loss activates CUP1 and HIS3 promoters to fully induced levels in the yeast Saccharomyces cerevisiae. Mol Cell Biol. 1992 Apr;12(4):1621–1629. doi: 10.1128/mcb.12.4.1621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Eisenmann D. M., Dollard C., Winston F. SPT15, the gene encoding the yeast TATA binding factor TFIID, is required for normal transcription initiation in vivo. Cell. 1989 Sep 22;58(6):1183–1191. doi: 10.1016/0092-8674(89)90516-3. [DOI] [PubMed] [Google Scholar]
  11. Errede B., Ammerer G. STE12, a protein involved in cell-type-specific transcription and signal transduction in yeast, is part of protein-DNA complexes. Genes Dev. 1989 Sep;3(9):1349–1361. doi: 10.1101/gad.3.9.1349. [DOI] [PubMed] [Google Scholar]
  12. Errede B., Company M., Ferchak J. D., Hutchison C. A., 3rd, Yarnell W. S. Activation regions in a yeast transposon have homology to mating type control sequences and to mammalian enhancers. Proc Natl Acad Sci U S A. 1985 Aug;82(16):5423–5427. doi: 10.1073/pnas.82.16.5423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Errede B., Company M., Hutchison C. A., 3rd Ty1 sequence with enhancer and mating-type-dependent regulatory activities. Mol Cell Biol. 1987 Jan;7(1):258–265. doi: 10.1128/mcb.7.1.258. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Farabaugh P., Liao X. B., Belcourt M., Zhao H., Kapakos J., Clare J. Enhancer and silencerlike sites within the transcribed portion of a Ty2 transposable element of Saccharomyces cerevisiae. Mol Cell Biol. 1989 Nov;9(11):4824–4834. doi: 10.1128/mcb.9.11.4824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fassler J. S., Winston F. Isolation and analysis of a novel class of suppressor of Ty insertion mutations in Saccharomyces cerevisiae. Genetics. 1988 Feb;118(2):203–212. doi: 10.1093/genetics/118.2.203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fulton A. M., Rathjen P. D., Kingsman S. M., Kingsman A. J. Upstream and downstream transcriptional control signals in the yeast retrotransposon, TY. Nucleic Acids Res. 1988 Jun 24;16(12):5439–5458. doi: 10.1093/nar/16.12.5439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Giaever G. N., Wang J. C. Supercoiling of intracellular DNA can occur in eukaryotic cells. Cell. 1988 Dec 2;55(5):849–856. doi: 10.1016/0092-8674(88)90140-7. [DOI] [PubMed] [Google Scholar]
  18. Goodbourn S. Negative regulation of transcriptional initiation in eukaryotes. Biochim Biophys Acta. 1990 Jun 1;1032(1):53–77. doi: 10.1016/0304-419x(90)90012-p. [DOI] [PubMed] [Google Scholar]
  19. Guarente L., Yocum R. R., Gifford P. A GAL10-CYC1 hybrid yeast promoter identifies the GAL4 regulatory region as an upstream site. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7410–7414. doi: 10.1073/pnas.79.23.7410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hansen L. J., Chalker D. L., Sandmeyer S. B. Ty3, a yeast retrotransposon associated with tRNA genes, has homology to animal retroviruses. Mol Cell Biol. 1988 Dec;8(12):5245–5256. doi: 10.1128/mcb.8.12.5245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hansen L. J., Sandmeyer S. B. Characterization of a transpositionally active Ty3 element and identification of the Ty3 integrase protein. J Virol. 1990 Jun;64(6):2599–2607. doi: 10.1128/jvi.64.6.2599-2607.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Happel A. M., Swanson M. S., Winston F. The SNF2, SNF5 and SNF6 genes are required for Ty transcription in Saccharomyces cerevisiae. Genetics. 1991 May;128(1):69–77. doi: 10.1093/genetics/128.1.69. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Levine M., Manley J. L. Transcriptional repression of eukaryotic promoters. Cell. 1989 Nov 3;59(3):405–408. doi: 10.1016/0092-8674(89)90024-x. [DOI] [PubMed] [Google Scholar]
  25. Liao X. B., Clare J. J., Farabaugh P. J. The upstream activation site of a Ty2 element of yeast is necessary but not sufficient to promote maximal transcription of the element. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8520–8524. doi: 10.1073/pnas.84.23.8520. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Liu L. F., Wang J. C. Supercoiling of the DNA template during transcription. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7024–7027. doi: 10.1073/pnas.84.20.7024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nagawa F., Fink G. R. The relationship between the "TATA" sequence and transcription initiation sites at the HIS4 gene of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8557–8561. doi: 10.1073/pnas.82.24.8557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Renkawitz R. Transcriptional repression in eukaryotes. Trends Genet. 1990 Jun;6(6):192–197. doi: 10.1016/0168-9525(90)90176-7. [DOI] [PubMed] [Google Scholar]
  29. Travers A. A. The reprogramming of transcriptional competence. Cell. 1992 May 15;69(4):573–575. doi: 10.1016/0092-8674(92)90218-2. [DOI] [PubMed] [Google Scholar]
  30. Tsao Y. P., Wu H. Y., Liu L. F. Transcription-driven supercoiling of DNA: direct biochemical evidence from in vitro studies. Cell. 1989 Jan 13;56(1):111–118. doi: 10.1016/0092-8674(89)90989-6. [DOI] [PubMed] [Google Scholar]
  31. Türkel S., Farabaugh P. J. Interspersion of an unusual GCN4 activation site with a complex transcriptional repression site in Ty2 elements of Saccharomyces cerevisiae. Mol Cell Biol. 1993 Apr;13(4):2091–2103. doi: 10.1128/mcb.13.4.2091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Voytas D. F., Boeke J. D. Yeast retrotransposon revealed. Nature. 1992 Aug 27;358(6389):717–717. doi: 10.1038/358717a0. [DOI] [PubMed] [Google Scholar]
  33. Winston F., Chaleff D. T., Valent B., Fink G. R. Mutations affecting Ty-mediated expression of the HIS4 gene of Saccharomyces cerevisiae. Genetics. 1984 Jun;107(2):179–197. doi: 10.1093/genetics/107.2.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Winston F., Dollard C., Malone E. A., Clare J., Kapakos J. G., Farabaugh P., Minehart P. L. Three genes are required for trans-activation of Ty transcription in yeast. Genetics. 1987 Apr;115(4):649–656. doi: 10.1093/genetics/115.4.649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Winston F., Durbin K. J., Fink G. R. The SPT3 gene is required for normal transcription of Ty elements in S. cerevisiae. Cell. 1984 Dec;39(3 Pt 2):675–682. doi: 10.1016/0092-8674(84)90474-4. [DOI] [PubMed] [Google Scholar]
  36. Wu H. Y., Shyy S. H., Wang J. C., Liu L. F. Transcription generates positively and negatively supercoiled domains in the template. Cell. 1988 May 6;53(3):433–440. doi: 10.1016/0092-8674(88)90163-8. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES