Deletion of the nuclear gene encoding the mitochondrial citrate transport protein from Saccharomyces cerevisiae

Ronald S. Kaplan, June A. Mayor, David Kakhniashvili, David A. Gremse, David O. Wood, David Nelson

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The nuclear gene encoding the mitochondrial citrate transport protein (i.e., CTP1) has been deleted from a haploid yeast strain. The stable yeast deletion strain was constructed by homologous recombination of the HIS3 gene at the CTP1 gene locus. Deletion of the CTP was confirmed by PCR. Immunoblot analysis provided the first quantitative estimate of the level of the CTP in wild-type yeast mitochondria and indicated the absence of expressed CTP in mitochondria isolated from the deletion strain. Deletion of CTP1 did not lead to a phenotype on any carbon source tested, indicating that CTP1 is not an essential gene. This suggests that either known alternative pathways are able to produce sufficient acetyl-CoA to support biosynthetic reactions, or there exists a second CTP gene. The ability of the deletion strain to serve as a host for the correct targeting and overexpression of a mutated CTP was then demonstrated. These studies provide a system which permits the use of sire-directed mutagenesis to examine both CTP targeting to mitochondria, as well as the molecular basis underlying CTP function. Moreover, this system will not only facilitate the study of the yeast CTP, but also CTPs expressed from the cDNAs of higher eukaryotes.

Original languageEnglish (US)
Pages (from-to)657-662
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume226
Issue number3
DOIs
StatePublished - Sep 24 1996

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Cytidine Triphosphate
Gene encoding
Mitochondrial Genes
Gene Deletion
Citric Acid
Yeast
Saccharomyces cerevisiae
Carrier Proteins
Mitochondria
Genes
Yeasts
Mutagenesis
Acetyl Coenzyme A
Haploidy
Homologous Recombination
Essential Genes
Eukaryota
Carbon
Complementary DNA
Phenotype

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Deletion of the nuclear gene encoding the mitochondrial citrate transport protein from Saccharomyces cerevisiae. / Kaplan, Ronald S.; Mayor, June A.; Kakhniashvili, David; Gremse, David A.; Wood, David O.; Nelson, David.

In: Biochemical and Biophysical Research Communications, Vol. 226, No. 3, 24.09.1996, p. 657-662.

Research output: Contribution to journalArticle

Kaplan, Ronald S. ; Mayor, June A. ; Kakhniashvili, David ; Gremse, David A. ; Wood, David O. ; Nelson, David. / Deletion of the nuclear gene encoding the mitochondrial citrate transport protein from Saccharomyces cerevisiae. In: Biochemical and Biophysical Research Communications. 1996 ; Vol. 226, No. 3. pp. 657-662.
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