Metabolic changes in Saccharomyces cerevisiae strains lacking citrate synthases

G. Kispál, M. Rosenkrantz, L. Guarente, P. Srere

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Abstract

The yeast, Saccharomyces cerevisiae, contains two citrate synthase isoenzymes, mitochondrial (CS1) and cytosolic (CS2). In this study, we have examined the metabolic consequences of the absence of CS1, CS2, and both isoenzymes in the respective mutant strains CS1-, CS2-, and CS1-CS2-. No significant differences were found in the growth rates of the parental, CS1-, or CS2- strains when grown in the single carbon sources galactose, glycerol, lactate, pyruvate, or glutamate. However, in nonfermentable carbon sources, the lag period in growth of CS1- was approximately 4 times that of the parental strain and the CS2- mutant. This difference was found even in glutamate. The CS1- mutant failed to grow on acetate in either complete or minimal liquid medium. Total cellular citrate concentration in the CS1- compared to the parental strain was higher when the cells were grown in lactate or pyruvate. On these same substrates, the malate concentration was 2-fold higher in the CS1- mutant when compared to the parental or CS2- strains. The production of 14CO2 by CS1- from [1-14C]acetate was 36% and that from [2-14C]acetate was 9.2% of the amount from the parental or CS2- strains. The 14CO2 production from [1-14C]glutamate was 28% and 20% in CS1- and CS1-CS2-, respectively, compared to the parental strain. Since these results are not easily explained solely by the absence of mitochondrial citrate synthase enzyme, we also determined the activity of some other enzymes of the citric acid cycle and electron transport chain. We found decreased activity of pyruvate dehydrogenase complex, α-ketoglutarate dehydrogenase complex, and aconitase, while the rest of the citric acid cycle enzymes and oxidative enzymes did not change significantly. The same changes in enzyme activities were found in two different yeast strains carrying the same citrate synthase mutations.

Original languageEnglish
Pages (from-to)11145-11149
Number of pages5
JournalJournal of Biological Chemistry
Volume263
Issue number23
Publication statusPublished - 1988

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Citrate (si)-Synthase
Yeast
Saccharomyces cerevisiae
Enzymes
Glutamic Acid
Acetates
Citric Acid Cycle
Pyruvic Acid
Isoenzymes
Lactic Acid
Carbon
Yeasts
Ketoglutarate Dehydrogenase Complex
Aconitate Hydratase
Pyruvate Dehydrogenase Complex
Growth
Electron Transport
Galactose
Citric Acid
Glycerol

ASJC Scopus subject areas

  • Biochemistry

Cite this

Metabolic changes in Saccharomyces cerevisiae strains lacking citrate synthases. / Kispál, G.; Rosenkrantz, M.; Guarente, L.; Srere, P.

In: Journal of Biological Chemistry, Vol. 263, No. 23, 1988, p. 11145-11149.

Research output: Contribution to journalArticle

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abstract = "The yeast, Saccharomyces cerevisiae, contains two citrate synthase isoenzymes, mitochondrial (CS1) and cytosolic (CS2). In this study, we have examined the metabolic consequences of the absence of CS1, CS2, and both isoenzymes in the respective mutant strains CS1-, CS2-, and CS1-CS2-. No significant differences were found in the growth rates of the parental, CS1-, or CS2- strains when grown in the single carbon sources galactose, glycerol, lactate, pyruvate, or glutamate. However, in nonfermentable carbon sources, the lag period in growth of CS1- was approximately 4 times that of the parental strain and the CS2- mutant. This difference was found even in glutamate. The CS1- mutant failed to grow on acetate in either complete or minimal liquid medium. Total cellular citrate concentration in the CS1- compared to the parental strain was higher when the cells were grown in lactate or pyruvate. On these same substrates, the malate concentration was 2-fold higher in the CS1- mutant when compared to the parental or CS2- strains. The production of 14CO2 by CS1- from [1-14C]acetate was 36{\%} and that from [2-14C]acetate was 9.2{\%} of the amount from the parental or CS2- strains. The 14CO2 production from [1-14C]glutamate was 28{\%} and 20{\%} in CS1- and CS1-CS2-, respectively, compared to the parental strain. Since these results are not easily explained solely by the absence of mitochondrial citrate synthase enzyme, we also determined the activity of some other enzymes of the citric acid cycle and electron transport chain. We found decreased activity of pyruvate dehydrogenase complex, α-ketoglutarate dehydrogenase complex, and aconitase, while the rest of the citric acid cycle enzymes and oxidative enzymes did not change significantly. The same changes in enzyme activities were found in two different yeast strains carrying the same citrate synthase mutations.",
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