Skip to main content
Log in

Effect of the pho85 Mutation on Catabolite Repression of the CIT1 Gene in Yeasts Saccharomyces cerevisiae

  • Published:
Russian Journal of Genetics Aims and scope Submit manuscript

Abstract

The Krebs cycle is one of the major metabolic pathways in a cell, which includes both catabolic and anabolic reactions. The first enzyme of the Krebs cycle, citrate synthase, catalyzes one of a few irreversible reactions of the cycle, citrate formation from acetyl-CoA and oxaloacetate. Expression of the CIT1 gene encoding the mitochondrial form of this enzyme inSaccharomyces cerevisiae is repressed on glucose- and glutamate-containing medium and activated on the raffinose-containing medium. In this work, the dependence of glucose repression of the CIT1 gene on the content of phosphate in the medium was studied. On the phosphate-deficient medium, the level of the CIT1 gene expression was increased twice. A low-molecular-weight (about 34 kDa) protein was identified and shown to interact with a region of the CIT1gene promoter (from –367 to –348 bp), which controls the glucose repression. The results obtained suggest that the Pho4 protein is involved in regulation of the CIT1gene expression on the glucose-containing and phosphate-deficient medium. Disruption of the PHO85 gene encoding phosphoprotein kinase (Pho4p is the substrate of this enzyme) leads to alleviation of glucose repression of the CIT1 gene. Thus, in yeast cells grown in the presence of glucose, the PHO85gene mediates downregulation of theCIT1expression.

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

  1. Klein, C.J.L., Olsson, L., and Nielsen, J., Glucose Control in Saccharomyces cerevisiae: The Role of MIG1 in Metabolic Functions, Microbiology, 1998, vol. 144, pp. 13-24.

    Google Scholar 

  2. Kotel'nikova, A.V. and Zvyagil'skaya, R.A., Biokhimiya drozhzhevykh mitokhondrii (Biochemistry of Yeast Mitochondria), Moscow: Nauka, 1973.

  3. Kim, K.-S., Rosenkrantz, M.S., and Guarente, L., Saccharomyces cerevisiae Contains Two Functional Citrate Synthase Genes, Mol. Cell. Biol., 1986, vol. 6, no. 6, pp. 1936-1942.

    Google Scholar 

  4. Rosenkrantz, M., Kell, C.S., Pennell, E.A., et al., Distinct Upstream Activation Regions for Glucose-Repressed and Derepressed Expression of the Yeast Citrate Synthase Gene, Curr. Genet., 1994, vol. 25, no. 3, pp. 185-195.

    Google Scholar 

  5. Samsonova, M.G., Smirnov, M.N., and Kozhin, S.A., A Genetic Biochemical Study of Acid Phosphatases of Yeast Saccharomyces cerevisiae: IX. Isolation and Analysis of Dominant Mutations Affecting Constitutive Synthesis of Acid Phosphatase II, Genetika (Moscow), 1980, vol. 16, no. 2, pp. 212-222.

    Google Scholar 

  6. Maniatis, T., Fritsch, E.F., and Sambrook, J., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor, New York: Cold Spring Harbor Lab., 1982.

    Google Scholar 

  7. Glover, D.M., DNA Cloning: A Practical Approach, Oxford: IRL, 1984.

    Google Scholar 

  8. Miller, J., Experiments in Molecular Genetics, Cold Spring Harbor, New York: Cold Spring Harbor Lab., 1976.

    Google Scholar 

  9. Toh-e, A., Ueda, Y., Kakimoto, S., et al., Isolation and Characterization of Acid Phosphatase Mutants in Saccharomyces cerevisiae, J. Bacteriol., 1973, vol. 113, pp. 727-738.

    Google Scholar 

  10. Urbakh, V.Yu., Biometricheskie metody (Biometrical Methods), Moscow: Nauka, 1964.

    Google Scholar 

  11. Bergman, L.W., Eisenberg, S., and Tye, B.-K., An Agarose Gel Electrophoresis Assay for the Detection of DNA-Binding Activities in Yeast Cell Extract, Methods Enzymol., 1987, vol. 195, pp. 528-537.

    Google Scholar 

  12. Bradford, M.M., A Rapid Sensitive Method for Quantitation of Microgram Quantities of Proteins Utilizing the Principle of Protein-Dye Binding, Anal. Biochem., 1981, vol. 72, pp. 247-251.

    Google Scholar 

  13. Hubscher, U., Double Replica Southwestern, Nucleic Acids Res., 1987, vol. 15, p. 5486.

    Google Scholar 

  14. Laemmli, U.K., Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4, Nature, 1970, vol. 15, pp. 680-685.

    Google Scholar 

  15. Pfeifer, K., Arcangioli, B., and Guarente, L., Yeast HAP1 Activator Competes with the Factor RC2 for Binding to the Upstream Activation Site UAS1 of the CYC1 Gene, Cell (Cambridge, Mass.), 1987, vol. 49, pp. 9-18.

    Google Scholar 

  16. Vogel, K., Horz, W., and Hinnen, A., The Two Positively Acting Regulatory Proteins PHO2 and PHO4 Physically Interact with PHO5 Upstream Activation Regions, Mol. Cell. Biol., 1989, vol. 9, pp. 2050-2057.

    Google Scholar 

  17. Liu, Z. and Butow, R.A., A Transcriptional Switch in the Expression of Yeast Tricarboxylic Acid Cycle Genes in Response to a Reduction or Loss of Respiratory Function, Mol. Cell. Biol., 1999, vol. 19, no. 10, pp. 6720-6728.

    Google Scholar 

  18. Shimizu, T., Toumoto, A., Ihara, K., et al., Crystal Structure of PHO4 bHLH Domain-DNA Complex: Flanking Base Recognition, EMBO J., 1997, vol. 16, pp. 4689-4697.

    Google Scholar 

  19. Jia, Y., Rothermel, J., Thornton, J., and Butow, R.A., A Basic Helix-Loop-Helix Zipper Transcription Complex Functions in a Signaling Pathway from Mitochondria to the Nucleus, Mol. Cell. Biol., 1997, vol. 17, pp. 1110-1117.

    Google Scholar 

  20. Shao, D., Creasy, C.L., and Bergman, L.W., A Cysteine Residue in Helix II of the HLH Domain Is Essential for Homodimerization of the Yeast Transcription Factor Pho4p, Nucleic Acids Res., 1998, vol. 26, pp. 710-714.

    Google Scholar 

  21. Kaffman, A., Hersskowitz, I., Tijan, R., et al., Phosphorylation of the Transcription Factor PHO4 by a Cyclin-CDK Complex, PHO80-PHO85, Science, 1994, vol. 263, pp. 1153-1156.

    Google Scholar 

  22. Timblin, B., Tatchell, K., and Bergman, L., Deletion of the Gene Encoding the Cyclin-Dependent Protein Kinase Pho85 Alters Glycogen Metabolism in Saccharomyces cerevisiae, Genetics, 1996, vol. 143, pp. 57-66.

    Google Scholar 

  23. Curtis Small, W., Brodeur, R.D., Sandor, A., et al., Enzymatic and Metabolic Studies on Retrograde Regulation Mutants of Yeast, Biochemistry, 1995, vol. 34, pp. 5569-5576.

    Google Scholar 

  24. Ogawa, N., De Risi, J., and Brown, P.O., New Components of a System for Phosphate Accumulation and Polyphosphate Metabolism in Saccharomyces cerevisiae Revealed by Genomic Expression Analysis, Mol. Biol. Cell, 2000, vol. 11, pp. 4309-4321.

    Google Scholar 

  25. Nishizawa, M., Tanabe, M., Yabuki, N., et al., Pho85 Kinase, a Yeast Cyclin-Dependent Kinase, Regulates the Expression of UGP1 Encoding UDP-Glucose Pyrophosphorylase, Yeast, 2001, vol. 18, no. 3, pp. 239-249.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Padkina, M.V., Tarasov, S.A., Karsten, S.L. et al. Effect of the pho85 Mutation on Catabolite Repression of the CIT1 Gene in Yeasts Saccharomyces cerevisiae . Russian Journal of Genetics 39, 604–609 (2003). https://doi.org/10.1023/A:1024489322873

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1024489322873

Keywords

Navigation