Title : Reversible transdominant inhibition of a metabolic pathway. In vivo evidence of interaction between two sequential tricarboxylic acid cycle enzymes in yeast.

Pub. Date : 2000 Apr 28

PMID : 10777592






4 Functional Relationships(s)
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1 We have isolated, in the Saccharomyces cerevisiae gene encoding the tricarboxylic acid cycle enzyme citrate synthase Cit1p, an "assembly mutation," i.e. a mutation that causes a tricarboxylic acid cycle deficiency without affecting the citrate synthase activity. Tricarboxylic Acids citrate (Si)-synthase CIT1 Saccharomyces cerevisiae S288C
2 We have isolated, in the Saccharomyces cerevisiae gene encoding the tricarboxylic acid cycle enzyme citrate synthase Cit1p, an "assembly mutation," i.e. a mutation that causes a tricarboxylic acid cycle deficiency without affecting the citrate synthase activity. Tricarboxylic Acids citrate (Si)-synthase CIT1 Saccharomyces cerevisiae S288C
3 We have shown that a 15-amino acid peptide from wild type Cit1p encompassing the mutation point inhibits the tricarboxylic acid cycle in a dominant manner, and that the inhibitory phenotype is overcome by a co-overexpression of Mdh1p, the mitochondrial malate dehydrogenase. Tricarboxylic Acids citrate (Si)-synthase CIT1 Saccharomyces cerevisiae S288C
4 These data provide the first direct in vivo evidence of interaction between two sequential tricarboxylic acid cycle enzymes, Cit1p and Mdh1p, and indicate that the characterization of assembly mutations by the reversible transdominant inhibition method may be a powerful way to study multienzyme complexes in their physiological context. Tricarboxylic Acids citrate (Si)-synthase CIT1 Saccharomyces cerevisiae S288C