Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1992 Nov;174(21):6831–6839. doi: 10.1128/jb.174.21.6831-6839.1992

Tripartite structure of the Saccharomyces cerevisiae arginase (CAR1) gene inducer-responsive upstream activation sequence.

M Viljoen 1, L Z Kovari 1, I A Kovari 1, H D Park 1, H J van Vuuren 1, T G Cooper 1
PMCID: PMC207359  PMID: 1400233

Abstract

Arginase (CAR1) gene expression in Saccharomyces cerevisiae is induced by arginine. The 5' regulatory region of CAR1 contains four separable regulatory elements--two inducer-independent upstream activation sequences (UASs) (UASC1 and UASC2), an inducer-dependent UAS (UASI), and an upstream repression sequence (URS1) which negatively regulates CAR1 and many other yeast genes. Here we demonstrate that three homologous DNA sequences originally reported to be present in the inducer-responsive UASI are in fact three exchangeable elements (UASI-A, UASI-B, and UASI-C). Although two of these elements, either the same or different ones, are required for transcriptional activation to occur, all three are required for maximal levels of induction. The elements operate in all orientations relative to one another and to the TATA sequence. All three UASI elements bind protein(s); protein binding does not require arginine or overproduction of any of the putative arginine pathway regulatory proteins. The UASI-protein complex was also observed even when extracts were derived from arg80/argRI or arg81/argRII deletion mutants. Similar sequences situated upstream of ARG5,6 and ARG3 and reported to negatively regulate their expression are able to functionally substitute for the CAR1 UASI elements and mediate reporter gene expression.

Full text

PDF
6833

Images in this article

Selected References

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

  1. Bossinger J., Cooper T. G. Molecular events associated with induction of arginase in Saccharomyces cerevisiae. J Bacteriol. 1977 Jul;131(1):163–173. doi: 10.1128/jb.131.1.163-173.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brindle P. K., Holland J. P., Willett C. E., Innis M. A., Holland M. J. Multiple factors bind the upstream activation sites of the yeast enolase genes ENO1 and ENO2: ABFI protein, like repressor activator protein RAP1, binds cis-acting sequences which modulate repression or activation of transcription. Mol Cell Biol. 1990 Sep;10(9):4872–4885. doi: 10.1128/mcb.10.9.4872. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cooper T. G., Kovari L., Sumrada R. A., Park H. D., Luche R. M., Kovari I. Nitrogen catabolite repression of arginase (CAR1) expression in Saccharomyces cerevisiae is derived from regulated inducer exclusion. J Bacteriol. 1992 Jan;174(1):48–55. doi: 10.1128/jb.174.1.48-55.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. De Rijcke M., Seneca S., Punyammalee B., Glansdorff N., Crabeel M. Characterization of the DNA target site for the yeast ARGR regulatory complex, a sequence able to mediate repression or induction by arginine. Mol Cell Biol. 1992 Jan;12(1):68–81. doi: 10.1128/mcb.12.1.68. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dubois E., Bercy J., Messenguy F. Characterization of two genes, ARGRI and ARGRIII required for specific regulation of arginine metabolism in yeast. Mol Gen Genet. 1987 Apr;207(1):142–148. doi: 10.1007/BF00331501. [DOI] [PubMed] [Google Scholar]
  6. Dubois E., Messenguy F. Isolation and characterization of the yeast ARGRII gene involved in regulating both anabolism and catabolism of arginine. Mol Gen Genet. 1985;198(2):283–289. doi: 10.1007/BF00383008. [DOI] [PubMed] [Google Scholar]
  7. Jacobs P., Jauniaux J. C., Grenson M. A cis-dominant regulatory mutation linked to the argB-argC gene cluster in Saccharomyces cerevisiae. J Mol Biol. 1980 Jun 5;139(4):691–704. doi: 10.1016/0022-2836(80)90055-8. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Keleher C. A., Goutte C., Johnson A. D. The yeast cell-type-specific repressor alpha 2 acts cooperatively with a non-cell-type-specific protein. Cell. 1988 Jun 17;53(6):927–936. doi: 10.1016/s0092-8674(88)90449-7. [DOI] [PubMed] [Google Scholar]
  10. Kovari L. Z., Cooper T. G. Participation of ABF-1 protein in expression of the Saccharomyces cerevisiae CAR1 gene. J Bacteriol. 1991 Oct;173(20):6332–6338. doi: 10.1128/jb.173.20.6332-6338.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kovari L., Sumrada R., Kovari I., Cooper T. G. Multiple positive and negative cis-acting elements mediate induced arginase (CAR1) gene expression in Saccharomyces cerevisiae. Mol Cell Biol. 1990 Oct;10(10):5087–5097. doi: 10.1128/mcb.10.10.5087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Luche R. M., Smart W. C., Cooper T. G. Purification of the heteromeric protein binding to the URS1 transcriptional repression site in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7412–7416. doi: 10.1073/pnas.89.16.7412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Luche R. M., Sumrada R., Cooper T. G. A cis-acting element present in multiple genes serves as a repressor protein binding site for the yeast CAR1 gene. Mol Cell Biol. 1990 Aug;10(8):3884–3895. doi: 10.1128/mcb.10.8.3884. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. MIDDELHOVEN W. J. THE PATHWAY OF ARGININE BREAKDOWN IN SACCHAROMYCES CEREVISIAE. Biochim Biophys Acta. 1964 Dec 9;93:650–652. doi: 10.1016/0304-4165(64)90349-6. [DOI] [PubMed] [Google Scholar]
  15. Messenguy F., Dubois E., Boonchird C. Determination of the DNA-binding sequences of ARGR proteins to arginine anabolic and catabolic promoters. Mol Cell Biol. 1991 May;11(5):2852–2863. doi: 10.1128/mcb.11.5.2852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Middelhoven W. J. Enzyme repression in the arginine pathway of Saccharomyces cerevisiae. Antonie Van Leeuwenhoek. 1969;35(2):215–226. doi: 10.1007/BF02219132. [DOI] [PubMed] [Google Scholar]
  17. Middelhoven W. J. Induction and repression of arginase and ornithine transaminase in baker's yeast. Antonie Van Leeuwenhoek. 1970;36(1):1–19. doi: 10.1007/BF02069003. [DOI] [PubMed] [Google Scholar]
  18. Middelhoven W. J. The derepression of arginase and of ornithine transaminase in nitrogen-starved baker's yeast. Biochim Biophys Acta. 1968 Mar 11;156(2):440–443. doi: 10.1016/0304-4165(68)90284-5. [DOI] [PubMed] [Google Scholar]
  19. Park H. D., Luche R. M., Cooper T. G. The yeast UME6 gene product is required for transcriptional repression mediated by the CAR1 URS1 repressor binding site. Nucleic Acids Res. 1992 Apr 25;20(8):1909–1915. doi: 10.1093/nar/20.8.1909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Qiu H. F., Dubois E., Broën P., Messenguy F. Functional analysis of ARGRI and ARGRIII regulatory proteins involved in the regulation of arginine metabolism in Saccharomyces cerevisiae. Mol Gen Genet. 1990 Jul;222(2-3):192–200. doi: 10.1007/BF00633817. [DOI] [PubMed] [Google Scholar]
  21. Qui H. F., Dubois E., Messenguy F. Dissection of the bifunctional ARGRII protein involved in the regulation of arginine anabolic and catabolic pathways. Mol Cell Biol. 1991 Apr;11(4):2169–2179. doi: 10.1128/mcb.11.4.2169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rai R., Genbauffe F. S., Sumrada R. A., Cooper T. G. Identification of sequences responsible for transcriptional activation of the allantoate permease gene in Saccharomyces cerevisiae. Mol Cell Biol. 1989 Feb;9(2):602–608. doi: 10.1128/mcb.9.2.602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Shore D., Stillman D. J., Brand A. H., Nasmyth K. A. Identification of silencer binding proteins from yeast: possible roles in SIR control and DNA replication. EMBO J. 1987 Feb;6(2):461–467. doi: 10.1002/j.1460-2075.1987.tb04776.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sumrada R. A., Cooper T. G. Isolation of the CAR1 gene from Saccharomyces cerevisiae and analysis of its expression. Mol Cell Biol. 1982 Dec;2(12):1514–1523. doi: 10.1128/mcb.2.12.1514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sumrada R. A., Cooper T. G. Point mutation generates constitutive expression of an inducible eukaryotic gene. Proc Natl Acad Sci U S A. 1985 Feb;82(3):643–647. doi: 10.1073/pnas.82.3.643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sumrada R. A., Cooper T. G. Ubiquitous upstream repression sequences control activation of the inducible arginase gene in yeast. Proc Natl Acad Sci U S A. 1987 Jun;84(12):3997–4001. doi: 10.1073/pnas.84.12.3997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Tabor S., Richardson C. C. DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4767–4771. doi: 10.1073/pnas.84.14.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Thiele D. J., Hamer D. H. Tandemly duplicated upstream control sequences mediate copper-induced transcription of the Saccharomyces cerevisiae copper-metallothionein gene. Mol Cell Biol. 1986 Apr;6(4):1158–1163. doi: 10.1128/mcb.6.4.1158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. West R. W., Jr, Chen S. M., Putz H., Butler G., Banerjee M. GAL1-GAL10 divergent promoter region of Saccharomyces cerevisiae contains negative control elements in addition to functionally separate and possibly overlapping upstream activating sequences. Genes Dev. 1987 Dec;1(10):1118–1131. doi: 10.1101/gad.1.10.1118. [DOI] [PubMed] [Google Scholar]
  30. Whitney P. A., Magasanik B. The induction of arginase in Saccharomyces cerevisiae. J Biol Chem. 1973 Sep 10;248(17):6197–6202. [PubMed] [Google Scholar]
  31. Wright C. F., Zitomer R. S. Point mutations implicate repeated sequences as essential elements of the CYC7 negative upstream site in Saccharomyces cerevisiae. Mol Cell Biol. 1985 Nov;5(11):2951–2958. doi: 10.1128/mcb.5.11.2951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yoo H. S., Cooper T. G. The DAL7 promoter consists of multiple elements that cooperatively mediate regulation of the gene's expression. Mol Cell Biol. 1989 Aug;9(8):3231–3243. doi: 10.1128/mcb.9.8.3231. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES