Yeast mutants of glucose metabolism with defects in the coordinate regulation of carbon assimilation

The enzymes of the glyoxylate cycle and gluconeogenesis are tightly regulat ed by transcriptional, posttranscriptional, and posttranslational mechanism s in Saccharomyces cerevisiae. We have previously identified four genes, AC N8, ACN9, ACN17, and ACN18, whose mutant phenotype includes two- to fourfol d elevated levels of enzymes of the glyoxylate cycle, gluconeogenesis, and acetyl-CoA metabolism. The affected enzymes are elevated on nonfermentable carbon sources but are still fully repressed by glucose. Catabolite inactiv ation of the cytosolic malate dehydrogenase is not affected in the mutants. Instead, the phenotype appeared to be manifested primarily at the level of transcription. The ACN8, ACN17, and ACN18 genes were isolated by functiona l complementation of the respective mutant's inability to utilize acetate a s a carbon and energy source, and these genes were shown to encode subunits of metabolic enzymes. ACN8 was identical to FBP1, which encodes the glucon eogenic enzyme, fructose 1,6-bisphosphatase, while ACN17 and ACN18 were ide ntical to the SDH2 and SDH4 genes, respectively, that encode subunits of th e respiratory chain and tricarboxylic acid cycle enzyme, succinate dehydrog enase. Mutants defective in other glyoxylate cycle and gluconeogenic enzyme s also display the elevated enzyme phenotype, indicating that the enzyme su perinduction is a general property of gluconeogenic dysfunction. Glucose 6- phosphate levels were diminished in the mutants, suggesting that endogenous glucose synthesis can regulate the expression of gluconeogenic enzymes. (C ) 1999 Academic Press.