Skip to main content
Log in

Mode of action of the gcr9 and cat3 mutations in restoring the ability of Saccharomyces cerevisiae tps1 mutants to grow on glucose

  • Original Paper
  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Abstract

Mutations in the TPS1 gene, which encodes trehalose-6-P synthase, cause a glucose-negative phenotype in Saccharomyces cerevisiae. Antimycin A or disruption of the QCR9 gene, which encodes one subunit of the cytochrome bc 1 complex, restore the ability to grow in glucose-containing media. Under these conditions the cell excreted a large amount of glycerol, corresponding to about 20% of the glucose taken up. Suppression appears to be achieved by diversion of accumulated glycolytic intermediates to the production of glycerol, thereby providing NAD+ and phosphate for the glyceraldehyde-3-P dehydrogenase reaction. Analysis of the mutation scil-1, which also suppresses the glucose-negative phenotype of tps1 mutants, showed that glucose transport was decreased in scil-1 mutants. The gene SCI1 was cloned and its nucleotide sequence revealed it to be identical to CAT3/SNF4. The suppression mediated by scil-1 is attributable to a decrease in glycolytic flux.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Albertyn J, Hohmann S, Thevelein JM, Prior B (1994) GPD1, which encodes glycerol-3-phosphate dehydrogenase, is essential for growth under osmotic stress in Saccharomyces cerevisiae, and its expression is regulated by the high osmolarity glycerol response pathway. Mol Cell Biol 14:4135–4144

    Google Scholar 

  • Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1994) Current protocols in molecular biology. Greene Publishing Associates — J. Wiley and Sons, New York

    Google Scholar 

  • Bergmeyer, HU (1983) UV methods with hexokinase and glucose-6-phosphate dehydrogenase. In: Kunst A, Draeger B, Ziegenhorn J (eds) Methods of enzymatic analysis, vol 6. Verlag Chemie, Weinheim, Germany, pp 163–172

    Google Scholar 

  • Bisson LF (1988) High affinity glucose transport in Saccharomyces cerevisiae is under general glucose repression control. J Bacteriol 170:4838–4845

    Google Scholar 

  • Bisson LF, Coons D, Kruckeberg AL, Lewis DA (1993) Yeast sugar transporters. Crit Rev Biochem Mol Biol 28:259–308

    Google Scholar 

  • Blázquez MA, Gancedo C (1994) Identification of extragenic suppressors of the cif1 mutation in Saccharomyces cerevisiae. Curr Genet 25:89–94

    Google Scholar 

  • Blázquez MA, Lagunas R, Gancedo C, Gancedo JM (1993) Trehalose-6-phosphate: a new regulator of yeast glycolysis that inhibits hexokinases. FEBS Lett 329:51–54

    Google Scholar 

  • Blázquez MA, Stucka R, Feldmann H, Gancedo C (1994) Trehalose-6-P synthase is dispensable for growth on glucose but not for spore germination in Schizosaccharomyces pombe. J. Bacteriol 176:3895–3902

    Google Scholar 

  • Broach JR, Strathern JN, Hicks JB (1979) Transformation in yeast: development of a hybrid cloning vector and isolation of the CAN1 gene. Gene 8:121–123

    Google Scholar 

  • Celenza JL, Eng FJ, Carlson M (1989) Molecular analysis of the SFF4 gene of Saccharomyces cerevisiae: evidence for physical association of the SNF4 protein with the SNF1 protein kinase. Mol Cell Biol 9:5045–5054

    Google Scholar 

  • De Vries S, Marres CAM (1987) The mitochondrial respiratory chain of yeast. Structure and biosynthesis and the role in cellular metabolism. Biochim Biophys Acta 895:205–239

    Google Scholar 

  • Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395

    Google Scholar 

  • Doi A, Doi K (1993) Correct end of the ORF for the CDC20 gene of Saccharomyces cerevisiae. EMBL Acc. No. D16506

  • Entian K-D, Zimmermann FK (1982) New genes involved in catabolite repression and derepression in the yeast Saccharomyces cerevisiae. J Bacteriol 151:1123–1128

    Google Scholar 

  • François J, Neves MJ, Hers HG (1991) The control of trehalose biosynthesis in Saccharomyces cerevisiae: evidence for a catabolite inactivation and repression of trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase. Yeast 7:575–587

    Google Scholar 

  • Gamo FJ (1995) Aislamiento y caracterizatión de mutaciones que suprimen el efecto tóxico de la glucosa en mutantes glicolíticos de Saccharomyces cerevisiae. Ph.D. Thesis. Universidad Autónoma de Madrid

  • Gamo FJ, Lafuente MJ, Gancedo C (1994) The mutation DGT1-1 decreases glucose transport and alleviates carbon catabolite repression in Saccharomyces cerevisiae. J Bacteriol 176:7423–7429

    Google Scholar 

  • Gancedo JM (1992) Carbon catabolite repression in yeast. Eur J Biochem 206:297–393

    Google Scholar 

  • González MI, Stucka R, Blázquez MA, Feldmann H, Gancedo C (1992) Molecular cloning of CIF1, a yeast gene necessary for growth on glucose. Yeast 8:183–192

    Google Scholar 

  • Hill JE, Myers AM, Koerner TJ, Tzagoloff A (1986) Yeast/E. coli shuttle vectors with multiple unique restriction sites. Yeast 2:163–167

    Google Scholar 

  • Hohmann S, Huse K, Valentin E, Mbonyi K, Thevelein JM, Zimmermann FK (1992) Glucose-induced regulatory defects in the Saccharomyces cerevisiae byp1 growth initiation mutant and identification of MIG1 as a partial suppressor. J Bacteriol 174:4183–4188

    Google Scholar 

  • Hohmann S, Neves MJ, de Koning W, Alijo R, Ramos J, Thevelein JM (1993) The growth and signalling defects of the ggs1 (fdp1/byp1) deletion mutant on glucose are suppressed by a deletion of the gene encoding hexokinase PII. Curr Genet 23:281–289

    Google Scholar 

  • Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168

    Google Scholar 

  • Johnson M (1941) The role of aerobic phosphorylation in the Pasteur effect. Science 94:200–202

    Google Scholar 

  • Koerner TJ, Hill J, Tzagoloff A (1985) Cloning and characterization of the yeast nuclear gene for subunit 5 of cytochrome oxidase. J Biol Chem 260:9513–9515

    Google Scholar 

  • Lagunas R (1986) Misconceptions about the energy metabolism of Saccharomyces cerevisiae. Yeast 2:222–228

    Google Scholar 

  • Luyten K, Albertyn J, Skibbe WF, Prior BA, Ramos J, Thevelein JM, Hohmann S (1995) FPS1, a yeast member of the MIP family of channel proteins, is a facilitator for glycerol uptake and efflux and is inactive under osmotic stress. EMBO J 14:1360–1371

    Google Scholar 

  • Lynen F (1941) Über den aeroben Phosphatbedarf der Hefe. Ein Beitrag zur Kenntnis der Pasteurschen Reaktion. Justus Liebig Ann Chem 546:120–141

    Google Scholar 

  • Navon G, Shulman RG, Yamame T, Eccleshall TR, Lam KB, Baronofsky JJ, Marmur J (1979) Phosphorus-31 nuclear magnetic resonance studies of wild type and glycolytic pathway mutants of Saccharomyces cerevisiae. Biochemistry 18:4487–4499

    Google Scholar 

  • Neigeborn L, Carlson M (1984) Genes affecting the regulation of SUC2 gene expression by glucose repression in Saccharomyces cerevisiae. Genetics 108:845–858

    Google Scholar 

  • Özcan S, Johnston M (1985) Three different regulatory mechanisms enable yeast hexose transporter (HXT) genes to be induced by different levels of glucose. Mol Cell Biol 15:1564–1572

    Google Scholar 

  • Phillips JD, Schmitt ME, Brown TA, Beckmann JD, Trumpower BL (1990) Isolation and characterization of QCR9, a nuclear gene encoding the 7.3 kDa subunit 9 of the Saccharomyces cerevisiae ubiquinol-cytochrome c oxidoreductase complex. J Biol Chem 265:20813–20821

    Google Scholar 

  • Robzyck C, Kassir Y (1992) A simple and highly efficient procedure for rescuing autonomous plasmids from yeast. Nucleic Acid Res 20:3790

    Google Scholar 

  • Rothstein R (1985) Cloning in yeast In: Glover DM (eds) DNA cloning: a practical approach, vol 2. IRL Press, Oxford-Washington, pp 45–65

    Google Scholar 

  • Rothstein R (1991) Targeting, disruption, replacement, and allele rescue: integrative DNA transformation in yeast. Methods Enzymol 194:281–301

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Schüller HJ, Entian KD (1988) Molecular characterization of the yeast regulatory gene CAT3 necessary for glucose derepression and nuclear localization of its product. Gene 67:247–257

    Google Scholar 

  • Sethi N, Monteagudo MC, Koshland D, Hogan E, Burke DJ (1991) The CDC20 gene product of Saccharomyces cerevisiae, a β-transducin homolog, is required for a subset of microtubule-dependent cellular processes. Mol Cell Biol 11:5592–5602

    Google Scholar 

  • Sikorski RS, Hieter P (1989) A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122:19–27

    Google Scholar 

  • Thevelein JM, Hohmann S (1995) Trehalose synthase: guard to the gate of glycolysis in yeast? Trends Biochem Sci 20:3–10

    Google Scholar 

  • Van Aelst L, Hohmann S, Bulaya B, de Koning W, Sierkstra L, Neves MJ, Luyten K, Alijo R, Ramos J, Coccetti P, Martegani E, Magalhaes-Rocha M, Lopes Brandao R, Van Dijck P, Vanhalewyn M, Durnez P, Jans AWH, Thevelein JM (1993) Molecular cloning of a gene involved in glucose sensing in the yeast Saccharomyces cerevisiae. Mol Microbiol 8:927–943

    Google Scholar 

  • Van Aelst L, Hohmann S, Zimmermann FK, Jans AWH, Thevelein JM (1991) A yeast homologue of the bovine lens fibre MIP gene family complements the growth defect of a Saccharomyces cerevisiae mutant on fermentable sugars but not its defect in glucose-induced RAS-mediated cAMP signalling. EMBO J 10:2095–2104

    Google Scholar 

  • Van de Poll KW, Kerkenaar A, Schamhart DHJ (1974) Isolation of a regulatory mutant of fructose-1,6-diphosphatase in Saccharomyces cerevisiae. J Bacteriol 117:965–970

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by C. P. Hollenberg

This paper is dedicated to Professor Friedrich K. Zimmermann on the occasion of his sixtieth birthday

Rights and permissions

Reprints and permissions

About this article

Cite this article

Blázquez, M.A., Gancedo, C. Mode of action of the gcr9 and cat3 mutations in restoring the ability of Saccharomyces cerevisiae tps1 mutants to grow on glucose. Molec. Gen. Genet. 249, 655–664 (1995). https://doi.org/10.1007/BF00418035

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00418035

Key words

Navigation