Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1996 Jan 15;15(2):363–374.

Genetic analysis of glucose regulation in saccharomyces cerevisiae: control of transcription versus mRNA turnover.

G P Cereghino 1, I E Scheffler 1
PMCID: PMC449951  PMID: 8617211

Abstract

A major determinant of the steady-state level of the mRNA encoding the iron protein (Ip) subunit of succinate dehydrogenase of yeast is its rate of turnover. This mRNA is significantly more stable in glycerol than in glucose media. Many other genes, for example, SUC2, that are repressed in the presence of glucose are believed to be controlled at the level of transcription. The present study elucidates differences in the regulatory mechanisms by which glucose controls the transcription and turnover of the SUC2 and Ip mRNAs. The signaling pathway for glucose repression at the transcriptional level has been associated with a number of gene products linking glucose uptake with nuclear events. We have investigated whether the same genes are involved in the control of Ip mRNA stability. Phosphorylation of glucose or fructose is critical in triggering the transcript's degradation, but any hexokinase will do. Of the other known genes examined, most, with the exception of REG1, are not involved in determining the differential stability of the Ip transcript. Finally, our results indicate that differential stability on different carbon sources also plays a role in determining the steady-state level of the SUC2 mRNA. Thus, glucose repression includes both transcriptional and post-transcriptional mechanisms.

Full text

PDF
363

Images in this article

Selected References

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

  1. Beelman C. A., Parker R. Degradation of mRNA in eukaryotes. Cell. 1995 Apr 21;81(2):179–183. doi: 10.1016/0092-8674(95)90326-7. [DOI] [PubMed] [Google Scholar]
  2. Bisson L. F., Coons D. M., Kruckeberg A. L., Lewis D. A. Yeast sugar transporters. Crit Rev Biochem Mol Biol. 1993;28(4):259–308. doi: 10.3109/10409239309078437. [DOI] [PubMed] [Google Scholar]
  3. Bisson L. F., Fraenkel D. G. Involvement of kinases in glucose and fructose uptake by Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1730–1734. doi: 10.1073/pnas.80.6.1730. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bisson L. F. High-affinity glucose transport in Saccharomyces cerevisiae is under general glucose repression control. J Bacteriol. 1988 Oct;170(10):4838–4845. doi: 10.1128/jb.170.10.4838-4845.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Carlson M., Osmond B. C., Botstein D. Mutants of yeast defective in sucrose utilization. Genetics. 1981 May;98(1):25–40. doi: 10.1093/genetics/98.1.25. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cereghino G. P., Atencio D. P., Saghbini M., Beiner J., Scheffler I. E. Glucose-dependent turnover of the mRNAs encoding succinate dehydrogenase peptides in Saccharomyces cerevisiae: sequence elements in the 5' untranslated region of the Ip mRNA play a dominant role. Mol Biol Cell. 1995 Sep;6(9):1125–1143. doi: 10.1091/mbc.6.9.1125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Elble R. A simple and efficient procedure for transformation of yeasts. Biotechniques. 1992 Jul;13(1):18–20. [PubMed] [Google Scholar]
  8. Entian K. D., Hilberg F., Opitz H., Mecke D. Cloning of hexokinase structural genes from Saccharomyces cerevisiae mutants with regulatory mutations responsible for glucose repression. Mol Cell Biol. 1985 Nov;5(11):3035–3040. doi: 10.1128/mcb.5.11.3035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Erickson J. R., Johnston M. Genetic and molecular characterization of GAL83: its interaction and similarities with other genes involved in glucose repression in Saccharomyces cerevisiae. Genetics. 1993 Nov;135(3):655–664. doi: 10.1093/genetics/135.3.655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Erickson J. R., Johnston M. Suppressors reveal two classes of glucose repression genes in the yeast Saccharomyces cerevisiae. Genetics. 1994 Apr;136(4):1271–1278. doi: 10.1093/genetics/136.4.1271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Estruch F., Carlson M. SNF6 encodes a nuclear protein that is required for expression of many genes in Saccharomyces cerevisiae. Mol Cell Biol. 1990 Jun;10(6):2544–2553. doi: 10.1128/mcb.10.6.2544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Estruch F., Treitel M. A., Yang X., Carlson M. N-terminal mutations modulate yeast SNF1 protein kinase function. Genetics. 1992 Nov;132(3):639–650. doi: 10.1093/genetics/132.3.639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Federoff H. J., Eccleshall T. R., Marmur J. Carbon catabolite repression of maltase synthesis in Saccharomyces carlsbergensis. J Bacteriol. 1983 Oct;156(1):301–307. doi: 10.1128/jb.156.1.301-307.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Flick J. S., Johnston M. GRR1 of Saccharomyces cerevisiae is required for glucose repression and encodes a protein with leucine-rich repeats. Mol Cell Biol. 1991 Oct;11(10):5101–5112. doi: 10.1128/mcb.11.10.5101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fraenkel D. G. Genetics and intermediary metabolism. Annu Rev Genet. 1992;26:159–177. doi: 10.1146/annurev.ge.26.120192.001111. [DOI] [PubMed] [Google Scholar]
  16. François J., Van Schaftingen E., Hers H. G. The mechanism by which glucose increases fructose 2,6-bisphosphate concentration in Saccharomyces cerevisiae. A cyclic-AMP-dependent activation of phosphofructokinase 2. Eur J Biochem. 1984 Nov 15;145(1):187–193. doi: 10.1111/j.1432-1033.1984.tb08539.x. [DOI] [PubMed] [Google Scholar]
  17. Hahn S., Guarente L. Yeast HAP2 and HAP3: transcriptional activators in a heteromeric complex. Science. 1988 Apr 15;240(4850):317–321. doi: 10.1126/science.2832951. [DOI] [PubMed] [Google Scholar]
  18. Herrick D., Parker R., Jacobson A. Identification and comparison of stable and unstable mRNAs in Saccharomyces cerevisiae. Mol Cell Biol. 1990 May;10(5):2269–2284. doi: 10.1128/mcb.10.5.2269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Lang J. M., Cirillo V. P. Glucose transport in a kinaseless Saccharomyces cerevisiae mutant. J Bacteriol. 1987 Jul;169(7):2932–2937. doi: 10.1128/jb.169.7.2932-2937.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Laurent B. C., Carlson M. Yeast SNF2/SWI2, SNF5, and SNF6 proteins function coordinately with the gene-specific transcriptional activators GAL4 and Bicoid. Genes Dev. 1992 Sep;6(9):1707–1715. doi: 10.1101/gad.6.9.1707. [DOI] [PubMed] [Google Scholar]
  22. Lederer B., Vissers S., Van Schaftingen E., Hers H. G. Fructose 2,6-bisphosphate in yeast. Biochem Biophys Res Commun. 1981 Dec 31;103(4):1281–1287. doi: 10.1016/0006-291x(81)90261-8. [DOI] [PubMed] [Google Scholar]
  23. Lobo Z., Maitra P. K. Resistance to 2-deoxyglucose in yeast: a direct selection of mutants lacking glucose-phosphorylating enzymes. Mol Gen Genet. 1977 Dec 9;157(3):297–300. doi: 10.1007/BF00268666. [DOI] [PubMed] [Google Scholar]
  24. Lombardo A., Cereghino G. P., Scheffler I. E. Control of mRNA turnover as a mechanism of glucose repression in Saccharomyces cerevisiae. Mol Cell Biol. 1992 Jul;12(7):2941–2948. doi: 10.1128/mcb.12.7.2941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ma H., Bloom L. M., Walsh C. T., Botstein D. The residual enzymatic phosphorylation activity of hexokinase II mutants is correlated with glucose repression in Saccharomyces cerevisiae. Mol Cell Biol. 1989 Dec;9(12):5643–5649. doi: 10.1128/mcb.9.12.5643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ma H., Bloom L. M., Zhu Z. M., Walsh C. T., Botstein D. Isolation and characterization of mutations in the HXK2 gene of Saccharomyces cerevisiae. Mol Cell Biol. 1989 Dec;9(12):5630–5642. doi: 10.1128/mcb.9.12.5630. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Neigeborn L., Carlson M. Mutations causing constitutive invertase synthesis in yeast: genetic interactions with snf mutations. Genetics. 1987 Feb;115(2):247–253. doi: 10.1093/genetics/115.2.247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Neigeborn L., Schwartzberg P., Reid R., Carlson M. Null mutations in the SNF3 gene of Saccharomyces cerevisiae cause a different phenotype than do previously isolated missense mutations. Mol Cell Biol. 1986 Nov;6(11):3569–3574. doi: 10.1128/mcb.6.11.3569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ng R., Abelson J. Isolation and sequence of the gene for actin in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3912–3916. doi: 10.1073/pnas.77.7.3912. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Niederacher D., Entian K. D. Characterization of Hex2 protein, a negative regulatory element necessary for glucose repression in yeast. Eur J Biochem. 1991 Sep 1;200(2):311–319. doi: 10.1111/j.1432-1033.1991.tb16187.x. [DOI] [PubMed] [Google Scholar]
  31. Nonet M., Scafe C., Sexton J., Young R. Eucaryotic RNA polymerase conditional mutant that rapidly ceases mRNA synthesis. Mol Cell Biol. 1987 May;7(5):1602–1611. doi: 10.1128/mcb.7.5.1602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Olesen J. T., Guarente L. The HAP2 subunit of yeast CCAAT transcriptional activator contains adjacent domains for subunit association and DNA recognition: model for the HAP2/3/4 complex. Genes Dev. 1990 Oct;4(10):1714–1729. doi: 10.1101/gad.4.10.1714. [DOI] [PubMed] [Google Scholar]
  33. Olesen J., Hahn S., Guarente L. Yeast HAP2 and HAP3 activators both bind to the CYC1 upstream activation site, UAS2, in an interdependent manner. Cell. 1987 Dec 24;51(6):953–961. doi: 10.1016/0092-8674(87)90582-4. [DOI] [PubMed] [Google Scholar]
  34. Ozcan S., Johnston M. Three different regulatory mechanisms enable yeast hexose transporter (HXT) genes to be induced by different levels of glucose. Mol Cell Biol. 1995 Mar;15(3):1564–1572. doi: 10.1128/mcb.15.3.1564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Ozcan S., Schulte F., Freidel K., Weber A., Ciriacy M. Glucose uptake and metabolism in grr1/cat80 mutants of Saccharomyces cerevisiae. Eur J Biochem. 1994 Sep 1;224(2):605–611. doi: 10.1111/j.1432-1033.1994.00605.x. [DOI] [PubMed] [Google Scholar]
  36. Peltz S. W., Brewer G., Bernstein P., Hart P. A., Ross J. Regulation of mRNA turnover in eukaryotic cells. Crit Rev Eukaryot Gene Expr. 1991;1(2):99–126. [PubMed] [Google Scholar]
  37. Pinkham J. L., Guarente L. Cloning and molecular analysis of the HAP2 locus: a global regulator of respiratory genes in Saccharomyces cerevisiae. Mol Cell Biol. 1985 Dec;5(12):3410–3416. doi: 10.1128/mcb.5.12.3410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Ronne H. Glucose repression in fungi. Trends Genet. 1995 Jan;11(1):12–17. doi: 10.1016/s0168-9525(00)88980-5. [DOI] [PubMed] [Google Scholar]
  39. Rose M., Albig W., Entian K. D. Glucose repression in Saccharomyces cerevisiae is directly associated with hexose phosphorylation by hexokinases PI and PII. Eur J Biochem. 1991 Aug 1;199(3):511–518. doi: 10.1111/j.1432-1033.1991.tb16149.x. [DOI] [PubMed] [Google Scholar]
  40. Schneider J. C., Guarente L. Regulation of the yeast CYT1 gene encoding cytochrome c1 by HAP1 and HAP2/3/4. Mol Cell Biol. 1991 Oct;11(10):4934–4942. doi: 10.1128/mcb.11.10.4934. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Sierkstra L. N., Silljé H. H., Verbakel J. M., Verrips C. T. The glucose-6-phosphate-isomerase reaction is essential for normal glucose repression in Saccharomyces cerevisiae. Eur J Biochem. 1993 May 15;214(1):121–127. doi: 10.1111/j.1432-1033.1993.tb17903.x. [DOI] [PubMed] [Google Scholar]
  42. Sierkstra L. N., Verbakel J. M., Verrips C. T. Analysis of transcription and translation of glycolytic enzymes in glucose-limited continuous cultures of Saccharomyces cerevisiae. J Gen Microbiol. 1992 Dec;138(12):2559–2566. doi: 10.1099/00221287-138-12-2559. [DOI] [PubMed] [Google Scholar]
  43. Surosky R. T., Strich R., Esposito R. E. The yeast UME5 gene regulates the stability of meiotic mRNAs in response to glucose. Mol Cell Biol. 1994 May;14(5):3446–3458. doi: 10.1128/mcb.14.5.3446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Szekely E., Montgomery D. L. Glucose represses transcription of Saccharomyces cerevisiae nuclear genes that encode mitochondrial components. Mol Cell Biol. 1984 May;4(5):939–946. doi: 10.1128/mcb.4.5.939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Thevelein J. M. Fermentable sugars and intracellular acidification as specific activators of the RAS-adenylate cyclase signalling pathway in yeast: the relationship to nutrient-induced cell cycle control. Mol Microbiol. 1991 Jun;5(6):1301–1307. doi: 10.1111/j.1365-2958.1991.tb00776.x. [DOI] [PubMed] [Google Scholar]
  46. Trawick J. D., Kraut N., Simon F. R., Poyton R. O. Regulation of yeast COX6 by the general transcription factor ABF1 and separate HAP2- and heme-responsive elements. Mol Cell Biol. 1992 May;12(5):2302–2314. doi: 10.1128/mcb.12.5.2302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Trumbly R. J. Cloning and characterization of the CYC8 gene mediating glucose repression in yeast. Gene. 1988 Dec 15;73(1):97–111. doi: 10.1016/0378-1119(88)90316-2. [DOI] [PubMed] [Google Scholar]
  48. Trumbly R. J. Glucose repression in the yeast Saccharomyces cerevisiae. Mol Microbiol. 1992 Jan;6(1):15–21. doi: 10.1111/j.1365-2958.1992.tb00832.x. [DOI] [PubMed] [Google Scholar]
  49. Trumbly R. J. Isolation of Saccharomyces cerevisiae mutants constitutive for invertase synthesis. J Bacteriol. 1986 Jun;166(3):1123–1127. doi: 10.1128/jb.166.3.1123-1127.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Ulery T. L., Jang S. H., Jaehning J. A. Glucose repression of yeast mitochondrial transcription: kinetics of derepression and role of nuclear genes. Mol Cell Biol. 1994 Feb;14(2):1160–1170. doi: 10.1128/mcb.14.2.1160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Vallier L. G., Coons D., Bisson L. F., Carlson M. Altered regulatory responses to glucose are associated with a glucose transport defect in grr1 mutants of Saccharomyces cerevisiae. Genetics. 1994 Apr;136(4):1279–1285. doi: 10.1093/genetics/136.4.1279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Williams F. E., Trumbly R. J. Characterization of TUP1, a mediator of glucose repression in Saccharomyces cerevisiae. Mol Cell Biol. 1990 Dec;10(12):6500–6511. doi: 10.1128/mcb.10.12.6500. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Williams F. E., Varanasi U., Trumbly R. J. The CYC8 and TUP1 proteins involved in glucose repression in Saccharomyces cerevisiae are associated in a protein complex. Mol Cell Biol. 1991 Jun;11(6):3307–3316. doi: 10.1128/mcb.11.6.3307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Winston F., Carlson M. Yeast SNF/SWI transcriptional activators and the SPT/SIN chromatin connection. Trends Genet. 1992 Nov;8(11):387–391. doi: 10.1016/0168-9525(92)90300-s. [DOI] [PubMed] [Google Scholar]
  55. Woods A., Munday M. R., Scott J., Yang X., Carlson M., Carling D. Yeast SNF1 is functionally related to mammalian AMP-activated protein kinase and regulates acetyl-CoA carboxylase in vivo. J Biol Chem. 1994 Jul 29;269(30):19509–19515. [PubMed] [Google Scholar]
  56. Zimmermann F. K., Scheel I. Mutants of Saccharomyces cerevisiae resistant to carbon catabolite repression. Mol Gen Genet. 1977 Jul 7;154(1):75–82. doi: 10.1007/BF00265579. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES