Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1979 Jul;139(1):152–160. doi: 10.1128/jb.139.1.152-160.1979

Physiological Effects of Seven Different Blocks in Glycolysis in Saccharomyces cerevisiae

Michael Ciriacy 1, Ingrid Breitenbach 1
PMCID: PMC216840  PMID: 378952

Abstract

Saccharomyces cerevisiae mutants unable to grow and ferment glucose have been isolated. Of 45 clones isolated, 25 had single enzyme defects of one of the following activities: phosphoglucose isomerase (pgi), phosphofructokinase (pfk), triosephosphate isomerase (tpi), phosphoglycerate kinase (pgk), phosphoglyceromutase (pgm), and pyruvate kinase (pyk). Phosphofructokinase activities in crude extracts of the pfk mutant were only 2% of the wild-type level. However, normal growth on glucose medium and normal fermentation of glucose suggested either that the mutant enzyme was considerably more active in vivo or, alternatively, that 2% residual activity was sufficient for normal glycolysis. All other mutants were moderately to strongly inhibited by glucose. Unusually high concentrations of glycolytic metabolites were observed before the reaction catalyzed by the enzyme which was absent in a given mutant strain when incubated on glucose. This confirmed at the cellular level the location of the defect as determined by enzyme assays. With adh (lacks all three alcohol dehydrogenase isozymes) and pgk mutants, accumulation of the typical levels of hexosephosphates was prevented when respiration was blocked with antimycin A. A typical feature of all glycolytic mutants described here was the rapid depletion of the intracellular adenosine 5′-triphosphate pool after transfer to glucose medium. No correlation of low or high levels of fructose-1,6-bisphosphate with the degree of catabolite repression and inactivation could be found. This observation does not support the concept that hexose metabolites are directly involved in these regulatory mechanisms in yeast.

Full text

PDF
152

Selected References

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

  1. Chance B. The nature of electron transfer and energy coupling reactions. FEBS Lett. 1972 Jun 1;23(1):3–20. doi: 10.1016/0014-5793(72)80272-2. [DOI] [PubMed] [Google Scholar]
  2. Ciriacy M. A yeast mutant with glucose-resistant formation of mitochondrial enzymes. Mol Gen Genet. 1978 Feb 27;159(3):329–335. doi: 10.1007/BF00268270. [DOI] [PubMed] [Google Scholar]
  3. Ciriacy M. Cis-dominant regulatory mutations affecting the formation of glucose-repressible alcohol dehydrogenase (ADHII) in Saccharomyces cerevisiae. Mol Gen Genet. 1976 Jun 15;145(3):327–333. doi: 10.1007/BF00325831. [DOI] [PubMed] [Google Scholar]
  4. Ciriacy M. Isolation and characterization of yeast mutants defective in intermediary carbon metabolism and in carbon catabolite derepression. Mol Gen Genet. 1977 Jul 20;154(2):213–220. doi: 10.1007/BF00330840. [DOI] [PubMed] [Google Scholar]
  5. Clifton D., Weinstock S. B., Fraenkel D. G. Glycolysis mutants in Saccharomyces cerevisiae. Genetics. 1978 Jan;88(1):1–11. doi: 10.1093/genetics/88.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. De Deken R. H. The Crabtree effect: a regulatory system in yeast. J Gen Microbiol. 1966 Aug;44(2):149–156. doi: 10.1099/00221287-44-2-149. [DOI] [PubMed] [Google Scholar]
  7. Gancedo J. M., Gancedo C. Concentrations of intermediary metabolites in yeast. Biochimie. 1973;55(2):205–211. doi: 10.1016/s0300-9084(73)80393-1. [DOI] [PubMed] [Google Scholar]
  8. Irani M. H., Maitra P. K. Properties of Escherichia coli mutants deficient in enzymes of glycolysis. J Bacteriol. 1977 Nov;132(2):398–410. doi: 10.1128/jb.132.2.398-410.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Lam K. B., Marmur J. Isolation and characterization of Saccharomyces cerevisiae glycolytic pathway mutants. J Bacteriol. 1977 May;130(2):746–749. doi: 10.1128/jb.130.2.746-749.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lobo Z., Maitra P. K. Genetics of yeast hexokinase. Genetics. 1977 Aug;86(4):727–744. doi: 10.1093/genetics/86.4.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lobo Z., Maitra P. K. Physiological role of glucose-phosphorylating enzymes in Saccharomyces cerevisiae. Arch Biochem Biophys. 1977 Aug;182(2):639–645. doi: 10.1016/0003-9861(77)90544-6. [DOI] [PubMed] [Google Scholar]
  13. Maitra P. K. Glucose and fructose metabolism in a phosphoglucoisomeraseless mutant of Saccharomyces cerevisiae. J Bacteriol. 1971 Sep;107(3):759–769. doi: 10.1128/jb.107.3.759-769.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Maitra P. K., Lobo Z. A kinetic study of glycolytic enzyme synthesis in yeast. J Biol Chem. 1971 Jan 25;246(2):475–488. [PubMed] [Google Scholar]
  15. Maitra P. K., Lobo Z. Pyruvate kinase mutants of Saccharomyces cerevisiae: biochemical and genetic characterisation. Mol Gen Genet. 1977 Apr 29;152(3):193–200. doi: 10.1007/BF00268817. [DOI] [PubMed] [Google Scholar]
  16. Polakis E. S., Bartley W. Changes in the enzyme activities of Saccharomyces cerevisiae during aerobic growth on different carbon sources. Biochem J. 1965 Oct;97(1):284–297. doi: 10.1042/bj0970284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sprague G. F., Jr Isolation and characterization of a Saccharomyces cerevisiae mutant deficient in pyruvate kinase activity. J Bacteriol. 1977 Apr;130(1):232–241. doi: 10.1128/jb.130.1.232-241.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Zimmermann F. K., Eaton N. R. Genetics of induction and catabolite repression of Maltese synthesis in Saccharomyces cerevisiae. Mol Gen Genet. 1974;134(3):261–272. doi: 10.1007/BF00267720. [DOI] [PubMed] [Google Scholar]

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

RESOURCES