Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1988 Feb;170(2):708–713. doi: 10.1128/jb.170.2.708-713.1988

Regulation of nitrogen assimilation in Saccharomyces cerevisiae: roles of the URE2 and GLN3 genes.

W E Courchesne 1, B Magasanik 1
PMCID: PMC210712  PMID: 2892826

Abstract

Mutations in the GLN3 gene prevented a normal increase in the NAD-glutamate dehydrogenase and glutamine synthetase levels in glutamate-grown Saccharomyces cerevisiae cells, whereas mutations in the URE2 gene resulted in high levels of these enzymes in glumate- and glutamine-grown cells. A ure2 gln3 double mutant had low levels of glutamate dehydrogenase and glutamine synthetase in cells grown on glutamate and glutamine; thus, gln3 mutations were epistatic to the ure2 mutations. The results suggest that the GLN3 product is capable of promoting increases in enzyme levels in the absence of a functional URE2 product and that the URE2 product antagonizes the GLN3 product. The URE2 and GLN3 genes were also found to regulate the level of arginase activity. This regulation is completely independent of the regulation of arginase by substrate induction. The activities of glutamate dehydrogenase, glutamine synthetase, and arginase were higher in cells grown on glutamate as the nitrogen source than they were in cells grown under a nitrogen-limiting condition. It had previously been shown that the levels of these enzymes can be increased by glutamine deprivation. We propose that the URE2-GLN3 system regulates enzyme synthesis, in response to glutamine and glutamate, to adjust the intracellular concentration of ammonia so as to maintain glutamine at the level required for optimal growth.

Full text

PDF
708

Selected References

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

  1. Brandriss M. C. Isolation and preliminary characterization of Saccharomyces cerevisiae proline auxotrophs. J Bacteriol. 1979 Jun;138(3):816–822. doi: 10.1128/jb.138.3.816-822.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brandriss M. C., Magasanik B. Genetics and physiology of proline utilization in Saccharomyces cerevisiae: mutation causing constitutive enzyme expression. J Bacteriol. 1979 Nov;140(2):504–507. doi: 10.1128/jb.140.2.504-507.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brandriss M. C., Magasanik B. Proline: an essential intermediate in arginine degradation in Saccharomyces cerevisiae. J Bacteriol. 1980 Sep;143(3):1403–1410. doi: 10.1128/jb.143.3.1403-1410.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Courchesne W. E., Magasanik B. Ammonia regulation of amino acid permeases in Saccharomyces cerevisiae. Mol Cell Biol. 1983 Apr;3(4):672–683. doi: 10.1128/mcb.3.4.672. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Deschamps J., Dubois E., Wiame J. M. L-Ornithine transaminase synthesis in Saccharomyces cerevisiae: regulation by inducer exclusion. Mol Gen Genet. 1979 Jul 24;174(3):225–232. doi: 10.1007/BF00267794. [DOI] [PubMed] [Google Scholar]
  6. Drillien R., Aigle M., Lacroute F. Yeast mutants pleiotropically impaired in the regulation of the two glutamate dehydrogenases. Biochem Biophys Res Commun. 1973 Jul 17;53(2):367–372. doi: 10.1016/0006-291x(73)90671-2. [DOI] [PubMed] [Google Scholar]
  7. Drillien R., Lacroute F. Ureidosuccinic acid uptake in yeast and some aspects of its regulation. J Bacteriol. 1972 Jan;109(1):203–208. doi: 10.1128/jb.109.1.203-208.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dubois E. L., Grenson M. Absence of involvement of glutamine synthetase and of NAD-linked glutamate dehydrogenase in the nitrogen catabolite repression of arginase and other enzymes in Saccharomyces cerevisiae. Biochem Biophys Res Commun. 1974 Sep 9;60(1):150–157. doi: 10.1016/0006-291x(74)90185-5. [DOI] [PubMed] [Google Scholar]
  9. Dubois E. L., Wiame J. M. Non specific induction of arginase in Saccharomyces cerevisiae. Biochimie. 1976;58(1-2):207–211. doi: 10.1016/s0300-9084(76)80371-9. [DOI] [PubMed] [Google Scholar]
  10. Dubois E., Vissers S., Grenson M., Wiame J. M. Glutamine and ammonia in nitrogen catabolite repression of Saccharomyces cerevisiae. Biochem Biophys Res Commun. 1977 Mar 21;75(2):233–239. doi: 10.1016/0006-291x(77)91033-6. [DOI] [PubMed] [Google Scholar]
  11. Grenson M., Dubois E., Piotrowska M., Drillien R., Aigle M. Ammonia assimilation in Saccharomyces cerevisiae as mediated by the two glutamate dehydrogenases. Evidence for the gdhA locus being a structural gene for the NADP-dependent glutamate dehydrogenase. Mol Gen Genet. 1974;128(1):73–85. doi: 10.1007/BF00267295. [DOI] [PubMed] [Google Scholar]
  12. HIERHOLZER G., HOLZER H. REPRESSION DER SYNTHESE VON DPN-ABHAENGIGER GLUTAMINSAEUREDEHYDROGENASE IN SACCHAROMYCES CEREVISIAE DURCH AMMONIUMIONEN. Biochem Z. 1963 Dec 3;339:175–185. [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Legrain C., Vissers S., Dubois E., Legrain M., Wiame J. M. Regulation of glutamine synthetase from Saccharomyces cerevisiae by repression, inactivation and proteolysis. Eur J Biochem. 1982 Apr;123(3):611–616. doi: 10.1111/j.1432-1033.1982.tb06576.x. [DOI] [PubMed] [Google Scholar]
  15. Lemoine Y., Dubois E., Wiame J. M. The regulation of urea amidolyase of Saccharomyces cerevisiae: mating type influence on a constitutivity mutation acting in cis. Mol Gen Genet. 1978 Nov 9;166(3):251–258. [PubMed] [Google Scholar]
  16. Mitchell A. P., Magasanik B. Purification and properties of glutamine synthetase from Saccharomyces cerevisiae. J Biol Chem. 1983 Jan 10;258(1):119–124. [PubMed] [Google Scholar]
  17. Mitchell A. P., Magasanik B. Regulation of glutamine-repressible gene products by the GLN3 function in Saccharomyces cerevisiae. Mol Cell Biol. 1984 Dec;4(12):2758–2766. doi: 10.1128/mcb.4.12.2758. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mitchell A. P., Magasanik B. Three regulatory systems control production of glutamine synthetase in Saccharomyces cerevisiae. Mol Cell Biol. 1984 Dec;4(12):2767–2773. doi: 10.1128/mcb.4.12.2767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mitchell A. P. The GLN1 locus of Saccharomyces cerevisiae encodes glutamine synthetase. Genetics. 1985 Oct;111(2):243–258. doi: 10.1093/genetics/111.2.243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sumrada R. A., Cooper T. G. Urea carboxylase and allophanate hydrolase are components of a multifunctional protein in yeast. J Biol Chem. 1982 Aug 10;257(15):9119–9127. [PubMed] [Google Scholar]
  21. 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]
  22. Wiame J. M., Grenson M., Arst H. N., Jr Nitrogen catabolite repression in yeasts and filamentous fungi. Adv Microb Physiol. 1985;26:1–88. doi: 10.1016/s0065-2911(08)60394-x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES