Assembly and function of a cytosolic form of NADH-specific isocitrate dehydrogenase in yeast

Wen Ning Zhao, Lee McAlister-Henn

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Mitochondrial NAD-dependent isocitrate dehydrogenase catalyzes a rate- limiting step in the tricarboxylic acid cycle. Yeast isocitrate dehydrogenase is an octomer composed of two subunits (IDH1 and IDH2) encoded by different genes and possessing independent mitochondrial targeting presequences. Oligonucleotide-directed mutagenesis was used to remove the presequences from each gene and from both genes carried on centromere-based expression plasmids. Effects on cellular localization were examined in a yeast strain containing chromosomal disruptions of IDH1 and IDH2 loci. Each subunit was found to be dependent upon its presequence for mitochondrial localization, and the subunits are independently imported into mitochondria under most growth conditions. Furthermore, an active holoenzyme can be assembled in the cytosol and this 'cytosolic' form of isocitrate dehydrogenase can reverse the acetate growth phenotype characteristic of the ΔIDH1/ΔIDH2 disruption strain, indicating functional replacement of the mitochondrial enzyme. However, transformants containing plasmids lacking either the IDH1 or IDH2 presequence coding regions were unexpectedly found to be capable of growth on acetate medium. Further investigation demonstrated that cellular localization of the IDH1 subunit can be biased by this stringent growth pressure.

Original languageEnglish (US)
Pages (from-to)10347-10352
Number of pages6
JournalJournal of Biological Chemistry
Volume271
Issue number17
DOIs
StatePublished - Apr 26 1996

Fingerprint

Isocitrate Dehydrogenase
NAD
Yeast
Yeasts
Genes
Growth
Acetates
Plasmids
Mutagenesis
Holoenzymes
Mitochondria
Citric Acid Cycle
Centromere
Site-Directed Mutagenesis
Oligonucleotides
Cytosol
Phenotype
Pressure
Enzymes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Assembly and function of a cytosolic form of NADH-specific isocitrate dehydrogenase in yeast. / Zhao, Wen Ning; McAlister-Henn, Lee.

In: Journal of Biological Chemistry, Vol. 271, No. 17, 26.04.1996, p. 10347-10352.

Research output: Contribution to journalArticle

Zhao, Wen Ning ; McAlister-Henn, Lee. / Assembly and function of a cytosolic form of NADH-specific isocitrate dehydrogenase in yeast. In: Journal of Biological Chemistry. 1996 ; Vol. 271, No. 17. pp. 10347-10352.
@article{4d82e998d0684dffb9ccc6ed87ea418b,
title = "Assembly and function of a cytosolic form of NADH-specific isocitrate dehydrogenase in yeast",
abstract = "Mitochondrial NAD-dependent isocitrate dehydrogenase catalyzes a rate- limiting step in the tricarboxylic acid cycle. Yeast isocitrate dehydrogenase is an octomer composed of two subunits (IDH1 and IDH2) encoded by different genes and possessing independent mitochondrial targeting presequences. Oligonucleotide-directed mutagenesis was used to remove the presequences from each gene and from both genes carried on centromere-based expression plasmids. Effects on cellular localization were examined in a yeast strain containing chromosomal disruptions of IDH1 and IDH2 loci. Each subunit was found to be dependent upon its presequence for mitochondrial localization, and the subunits are independently imported into mitochondria under most growth conditions. Furthermore, an active holoenzyme can be assembled in the cytosol and this 'cytosolic' form of isocitrate dehydrogenase can reverse the acetate growth phenotype characteristic of the ΔIDH1/ΔIDH2 disruption strain, indicating functional replacement of the mitochondrial enzyme. However, transformants containing plasmids lacking either the IDH1 or IDH2 presequence coding regions were unexpectedly found to be capable of growth on acetate medium. Further investigation demonstrated that cellular localization of the IDH1 subunit can be biased by this stringent growth pressure.",
author = "Zhao, {Wen Ning} and Lee McAlister-Henn",
year = "1996",
month = "4",
day = "26",
doi = "10.1074/jbc.271.17.10347",
language = "English (US)",
volume = "271",
pages = "10347--10352",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "17",

}

TY - JOUR

T1 - Assembly and function of a cytosolic form of NADH-specific isocitrate dehydrogenase in yeast

AU - Zhao, Wen Ning

AU - McAlister-Henn, Lee

PY - 1996/4/26

Y1 - 1996/4/26

N2 - Mitochondrial NAD-dependent isocitrate dehydrogenase catalyzes a rate- limiting step in the tricarboxylic acid cycle. Yeast isocitrate dehydrogenase is an octomer composed of two subunits (IDH1 and IDH2) encoded by different genes and possessing independent mitochondrial targeting presequences. Oligonucleotide-directed mutagenesis was used to remove the presequences from each gene and from both genes carried on centromere-based expression plasmids. Effects on cellular localization were examined in a yeast strain containing chromosomal disruptions of IDH1 and IDH2 loci. Each subunit was found to be dependent upon its presequence for mitochondrial localization, and the subunits are independently imported into mitochondria under most growth conditions. Furthermore, an active holoenzyme can be assembled in the cytosol and this 'cytosolic' form of isocitrate dehydrogenase can reverse the acetate growth phenotype characteristic of the ΔIDH1/ΔIDH2 disruption strain, indicating functional replacement of the mitochondrial enzyme. However, transformants containing plasmids lacking either the IDH1 or IDH2 presequence coding regions were unexpectedly found to be capable of growth on acetate medium. Further investigation demonstrated that cellular localization of the IDH1 subunit can be biased by this stringent growth pressure.

AB - Mitochondrial NAD-dependent isocitrate dehydrogenase catalyzes a rate- limiting step in the tricarboxylic acid cycle. Yeast isocitrate dehydrogenase is an octomer composed of two subunits (IDH1 and IDH2) encoded by different genes and possessing independent mitochondrial targeting presequences. Oligonucleotide-directed mutagenesis was used to remove the presequences from each gene and from both genes carried on centromere-based expression plasmids. Effects on cellular localization were examined in a yeast strain containing chromosomal disruptions of IDH1 and IDH2 loci. Each subunit was found to be dependent upon its presequence for mitochondrial localization, and the subunits are independently imported into mitochondria under most growth conditions. Furthermore, an active holoenzyme can be assembled in the cytosol and this 'cytosolic' form of isocitrate dehydrogenase can reverse the acetate growth phenotype characteristic of the ΔIDH1/ΔIDH2 disruption strain, indicating functional replacement of the mitochondrial enzyme. However, transformants containing plasmids lacking either the IDH1 or IDH2 presequence coding regions were unexpectedly found to be capable of growth on acetate medium. Further investigation demonstrated that cellular localization of the IDH1 subunit can be biased by this stringent growth pressure.

UR - http://www.scopus.com/inward/record.url?scp=0029971335&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029971335&partnerID=8YFLogxK

U2 - 10.1074/jbc.271.17.10347

DO - 10.1074/jbc.271.17.10347

M3 - Article

C2 - 8626605

AN - SCOPUS:0029971335

VL - 271

SP - 10347

EP - 10352

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 17

ER -