Extramitochondrial citrate synthase activity in bakers' yeast.

Rickey, T M; Lewin, Alfred S.

doi:10.1128/mcb.6.2.488

Cited by 33 publications

(29 citation statements)

References 29 publications

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“…Disruption of both genes was necessary to produce these phenotypes. Our results agree with a recent report that disruption of CITJ does not eliminate citrate synthase activity or produce citrate synthase-deficient growth phenotypes (30). We therefore conclude that citrate synthase in S. cerevisiae is encoded by the two loci CITJ and CIT2 and that the product of either gene can supply a level of citrate sufficient for biosynthesis of glutamate and for optimal growth on rich lactate medium.…”

Section: Methodssupporting

confidence: 82%

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Saccharomyces cerevisiae contains two functional citrate synthase genes.

Kim¹,

Rosenkrantz²,

Guarente³

1986

Mol. Cell. Biol.

137

123

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The tricarboxylic acid cycle occurs within the mitochondria of the yeast Saccharomyces cerevisiae. A nuclear gene encoding the tricarboxylic acid cycle enzyme citrate synthase has previously been isolated (M. Suissa, K. Suda, and G. Schatz, EMBO J. 3:1773-1781, 1984) and is referred to here as CIT1. We report here the isolation, by an immunological method, of a second nuclear gene encoding citrate synthase (CIT2). Disruption of both genes in the yeast genome was necessary to produce classical citrate synthase-deficient phenotypes: glutamate auxotrophy and poor growth on rich medium containing lactate, a nonfermentable carbon source. Therefore, the citrate synthase produced from either gene was sufficient for these metabolic roles. Transcription of both genes was maximally repressed in medium containing both glucose and glutamate. However, transcription of CIT1 but not of CIT2 was derepressed in medium containing a nonfermentable carbon source. The significance of the presence of two genes encoding citrate synthase in S. cerevisiae is discussed.

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Section: Methodssupporting

confidence: 82%

“…The predicted amino acid sequence of this product is highly homologous to the sequence of pig heart and Escherichia coli citrate synthases. Recently, it has been shown that disruption of this gene in the yeast genome severely reduces mitochondrial citrate synthase activity but does not reduce extramitochondrial activity (30).…”

mentioning

confidence: 99%

Saccharomyces cerevisiae contains two functional citrate synthase genes.

Kim¹,

Rosenkrantz²,

Guarente³

1986

Mol. Cell. Biol.

137

123

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“…Saccharomyces cerevisiae contains two forms of citrate synthase, which are encoded by distinct nuclear genes (14,20,21,25). DNA sequence analysis reveals that the two isozymes are similar in size and primary structure except at their amino termini.…”

mentioning

confidence: 99%

“…3). This antiserum does not react with the extramitochondrial isozyme (20). In addition to exposing the filter to X-ray film, we measured radioactivity directly for 5 h, using an Ambis radioanalytic imaging system.…”

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Citrate synthase encoded by the CIT2 gene of Saccharomyces cerevisiae is peroxisomal.

1990

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The product of the CIT2 gene has the tripeptide SKL at its carboxyl terminus. This amino acid sequence has been shown to act as a peroxisomal targeting signal in mammalian cells. We examined the subcellular site of this extramitochondrial citrate synthase. Cells of Saccharomyces cerevisiae were grown on oleate medium to induce peroxisome proliferation. A fraction containing membrane-enclosed vesicles and organelles was analyzed by sedimentation on density gradients. In wild-type cells, the major peak of citrate synthase activity was recovered in the mitochondrial fraction, but a second peak of activity cosedimented with peroxisomes. The peroxisomal activity, but not the mitochondrial activity, was inhibited by incubation at pH 8.1, a characteristic of the extramitochondrial citrate synthase encoded by the CIT2 gene. In a strain in which the CITI gene encoding mitochondrial citrate synthase had been disrupted, the major peak of citrate synthase activity was peroxisomal, and all of the activity was sensitive to incubation at pH 8.1. Yeast cells bearing a cit2 disruption were unable to mobilize stored lipids and did not form stable peroxisomes in oleate. We conclude that citrate synthase encoded by CIT2 is peroxisomal and participates in the glyoxylate cycle.

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The complete sequence of a 10·8kb fragment to the right of the chromosome III centromere of Saccharomyces cerevisiae

Biteau

Fremaux

Hébrard

et al. 1992

Yeast

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The complete nucleotide sequence of the D10H fragment (10850 bp) was determined. The D10H fragment is located on the right arm of chromosome III near the centromere and contains the SUF2 gene. Six open reading frames (ORFs) larger than 300 bp were found. One of them is the CIT2 gene encoding the cytoplasmic citrate synthase. The others are new putative genes and show no significant similarity with any known gene. In addition two tRNA genes (Asn and Pro) and a solo delta element were identified. Two ORFs were disrupted; no peculiar phenotype was observed.

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Extramitochondrial citrate synthase activity in bakers' yeast.

Cited by 33 publications

References 29 publications

Saccharomyces cerevisiae contains two functional citrate synthase genes.

Saccharomyces cerevisiae contains two functional citrate synthase genes.

Citrate synthase encoded by the CIT2 gene of Saccharomyces cerevisiae is peroxisomal.

The complete sequence of a 10·8kb fragment to the right of the chromosome III centromere of Saccharomyces cerevisiae

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