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 language | English (US) |
|---|---|
| Pages (from-to) | 10347-10352 |
| Number of pages | 6 |
| Journal | Journal of Biological Chemistry |
| Volume | 271 |
| Issue number | 17 |
| DOIs | |
| State | Published - Apr 26 1996 |
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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 journal › Article
}
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.
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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 -