Expression and gene disruption analysis of the isocitrate dehydrogenase family in yeast

Wen Ning Zhao, Lee McAlister-Henn

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Mammalian and yeast cells contain three isozymes of isocitrate dehydrogenase: mitochondrial NAD- and NADP-specific enzymes and a cytosolic NADP-specific enzyme. Independent metabolic functions of these enzymes in Saccharomyces cerevisiae were examined by analyses of expression and of phenotypes displayed by mutants containing all possible combinations of isozyme gene disruptions. All three isocitrate dehydrogenases are expressed at high levels with growth on nonfermentable carbon sources, whereas the mitochondrial NADP-specific enzyme constitutes the major cellular activity with growth on glucose. Distinct growth phenotypes are observed for mutants expressing a single isozyme, and expression of at least one isozyme is necessary for glutamate-independent growth. The NADP-specific tricarboxylic acid cycle isocitrate dehydrogenase from Escherichia coli was expressed in mitochondrial and cytosolic compartments of the yeast disruption mutants using plasmids carrying gene fusions of yeast promoters and a mitochondrial targeting presequence with the bacterial coding sequence. The bacterial enzyme is competent for restoration of NADP-specific functions in either compartment but does not compensate for function of the yeast NAD-specific tricarboxylic acid cycle enzyme.

Original languageEnglish (US)
Pages (from-to)7873-7878
Number of pages6
JournalBiochemistry
Volume35
Issue number24
DOIs
StatePublished - Jun 18 1996

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Isocitrate Dehydrogenase
Yeast
NADP
Genes
Yeasts
Gene Expression
Isoenzymes
Enzymes
Citric Acid Cycle
Growth
NAD
Phenotype
Gene Fusion
Escherichia coli
Restoration
Saccharomyces cerevisiae
Glutamic Acid
Plasmids
Fusion reactions
Carbon

ASJC Scopus subject areas

  • Biochemistry

Cite this

Expression and gene disruption analysis of the isocitrate dehydrogenase family in yeast. / Zhao, Wen Ning; McAlister-Henn, Lee.

In: Biochemistry, Vol. 35, No. 24, 18.06.1996, p. 7873-7878.

Research output: Contribution to journalArticle

Zhao, Wen Ning ; McAlister-Henn, Lee. / Expression and gene disruption analysis of the isocitrate dehydrogenase family in yeast. In: Biochemistry. 1996 ; Vol. 35, No. 24. pp. 7873-7878.
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