Skip to main content
SpringerLink
Log in

Identification and characterization of regulatory elements in the phosphoenolpyruvate carboxykinase gene PCK1 of Saccharomyces cerevisiae

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

Abstract

Phosphoenolpyruvate carboxykinase is a key enzyme in gluconeogenesis. The expression of the PCK1 gene in Saccharomyces cerevisiae is strictly regulated and dependent on the carbon source provided. Two upstream activation sites (UAS1PCK1 and UAS2PCK1) and one upstream repression site (URSPCK1) were localized by detailed deletion analysis. The efficacy of these three promoter elements when separated from each other was confirmed by investigations using heterologous promoter test plasmids. Activation mediated by UAS1PCK1 or UAS2PCK1 did not occur in the presence of glucose, indicating that these elements are essential for glucose derepression. The repressing effect caused by URSPCK1 was much stronger in glucose-grown cells than in ethanol-grown cells.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1987–1989) Current protocols in molecular biology. John Wiley and Sons, New York

    Google Scholar 

  • Burlini N, Morandi S, Pellegrini R, Tortora P, Guerritore A (1989) Studies on the degradative mechanism of phosphoenolpyruvate carboxykinase from the yeast Saccharomyces cerevisiae. Biochim Biophys Acta 1014:153–161

    Google Scholar 

  • Celenza JL, Carlson M (1986) A yeast gene that is essential for release from glucose repression encodes a protein kinase. Science 233:1175–1180

    Google Scholar 

  • Celenza JL, Eng FJ, Carlson M (1989) Molecular analysis of the SNF4 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 

  • Ciriacy M (1975) Genetics of alcohol dehydrogenase in Saccharomyces cerevisiae. Mutat Res 29:315–326

    Google Scholar 

  • Entian KD (1986) Glucose repression: a complex regulatory system in yeast. Microbiol Sci 3:366–371

    Google Scholar 

  • Entian KD, Barnett JA (1992) Regulation of sugar utilization by Saccharomyces cerevisiae. TIBS 17:506–510

    Google Scholar 

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

    Google Scholar 

  • Fernandez E, Fernandez M, Moreno F, Rodicio R (1993) Transcriptional regulation of the isocitrate lyase encoding gene in Saccharomyces cerevisiae. FEBS Lett 333:238–242

    Google Scholar 

  • Gancedo JH (1992) Carbon catabolite repression in yeast. Eur J Biochem 206:297–313

    Google Scholar 

  • Gancedo C, Schwerzmann N (1976) Inactivation by glucose of phosphoenolpyruvate carboxykinase from Saccharomyces cerevisiae. Arch Microbiol 109:221–225

    Google Scholar 

  • Guarente L (1983) Yeast promoters and lacZ fusions designed to study expression of cloned genes in yeast. Methods Enzymol 101:181–191

    Google Scholar 

  • Haarasilta S, Oura E (1975) On the activity and regulation of anaplerotic and gluconeogenetic enzymes during the growth process of baker's yeast - the biphasic growth. Eur J Biochem 52:1–7

    Google Scholar 

  • Klebe RJ, Harriss JV, Sharp D, Douglas MG (1983) A general method for polyethylenglycol-induced transformation of bacteria and yeast. Gene 25:333–341

    Google Scholar 

  • Luche RM, Sumrada R, Cooper TG (1990) A cis-acting element present in multiple genes serves as a repressor protein binding site for the yeast CAR1 gene. Mol Cell Biol 10:3884–3895

    Google Scholar 

  • Luche RM, Smart WC, Cooper TG (1992) Purification of the heteromeric protein binding to the URS1 transcriptional repression site in Saccharomyces cerevisiae. Proc Natl Acad Sci USA 89:7412–7416

    Google Scholar 

  • Mercado JJ, Gancedo JM (1992) Regulatory regions in the yeast FBP1 and PCK1 genes. FEBS Lett. 311:110–114

    Google Scholar 

  • Myers AM, Tzagoloff A, Kinney DM, Lusty CJ (1986) Yeast shuttle and integrative vectors with multiple cloning sites suitable for construction of lacZ fusions. Gene 45:299–310

    Google Scholar 

  • Nehlin IO, Ronne H (1990) Yeast Mig1 repressor is related to the mammalian early growth response and Wilm's tumour finger proteins. EMBO J 10:3373–3378

    Google Scholar 

  • Niederacher D, Entian KD (1987) Isolation and characterization of the regulatory HEX2 gene necessary for glucose repression in yeast. Mol Gen Genet 206:505–509

    Google Scholar 

  • Niederacher D, Schüller HJ, Grzesitza D, Gütlich H, Hauser HP, Wagner T, Entian KD (1992) Identification of UAS elements and binding proteins necessary for derepression of Saccharomyces cerevisiae fructose-1,6-bisphosphatase. Curr Genet 21:363–370

    Google Scholar 

  • Park HO, Craig EA (1991) Transcriptional regulation of a yeast HSP70 gene by heat shock factor and an upstream repression site-binding factor. Genes Dev 5:1299–1308

    Google Scholar 

  • Schöler A, Schüller HJ (1993) Structure and regulation of the isocitrate lyase gene ICL1 from the yeast Saccharomyces cerevisiae. Curr Genet 23:375–381

    Google Scholar 

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

    Google Scholar 

  • Schüller HJ, Hahn A, Tröster F, Schütz A, Schweizer E (1992) Coordinate genetic control of yeast fatty acid synthase genes FAS1 and FAS2 by an upstream activation site common to genes involved in membrane lipid biosynthesis. EMBO J 11:107–114

    Google Scholar 

  • Shore D, Nasmyth K (1987) Purification and cloning of a DNA-binding protein that binds to both silencer and activator elements. Cell 51:721–732

    Google Scholar 

  • Tautz D, Renz M (1983) An optimized freeze squeeze method for the recovery of DNA fragments from agarose gels. Anal Biochem 132:14–19

    Google Scholar 

  • Trumbly RJ (1992) Glucose repression in the yeast Saccharomyces cerevisiae. Mol Microbiol 6:15–21

    Google Scholar 

  • Valdés-Hevia MD (1989) Isolation and characterization of the gene encoding phosphoenolpyruvate carboxykinase from Saccharomyces cerevisiae. FEBS Lett 258:313–316

    Google Scholar 

  • Zamenhoff S (1957) Preparation and assay of deoxyribonucleic acids from animal tissue. Methods Enzymol 3:696–704

    Google Scholar 

  • Zimmermann FK (1977) Mutants of Saccharomyces cerevisiae resistant to carbon catabolite repression. Mol Gen Genet 154:75–82

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Mikrobiologie, Johann Wolfgang Goethe-Universität, Biozentrum Niederursel, Marie-Curie-Straße 9, D-60439, Frankfurt am Main, Germany

    M. Proft & D. Grzesitza

Authors
  1. M. Proft
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. D. Grzesitza
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by C. Hollenberg

Rights and permissions

Reprints and permissions

About this article

Cite this article

Proft, M., Grzesitza, D. Identification and characterization of regulatory elements in the phosphoenolpyruvate carboxykinase gene PCK1 of Saccharomyces cerevisiae . Molec. Gen. Genet. 246, 367–373 (1995). https://doi.org/10.1007/BF00288610

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00288610

Key words

Search

Navigation