Upstream region of the SUC2 gene confers regulated expression to a heterologous gene in Saccharomyces cerevisiae.
The SUC2 gene produces two differently regulated mRNAs that encode two forms of invertase. The 1.9-kilobase mRNA encoding secreted invertase is regulated by glucose (carbon catabolite) repression, and the 1.8-kilobase mRNA encoding intracellular invertase is synthesized constitutively. Previous work has shown that the 5' noncoding region between -650 and -418 is required for derepression of secreted invertase in response to glucose deprivation. We show here that this upstream region can confer glucose-repressible expression to a heterologous gene, a LEU2-lacZ gene fusion, that is not normally regulated by glucose repression. This expression was found to respond appropriately to mutations in trans-acting genes that affect regulation of SUC2 expression. Mutations in the SNF1 through SNF6 loci reduced derepression of beta-galactosidase, and a mutation at the SSN6 locus caused constitutive expression. These findings indicate that the SUC2 upstream region mediates the regulatory effects of these genes and suggest that regulation occurs at the level of transcription. In addition, the upstream region was partially active in the inverted orientation.
D007446:Chromosome Inversion; D004271:DNA, Fungal / Q000235:genetics; D005786:Gene Expression Regulation; D005800:Genes, Fungal; D005809:Genes, Regulator; D005818:Genetic Engineering; D005947:Glucose / Q000502:physiology; D006026:Glycoside Hydrolases / Q000235:genetics*; D009154:Mutation; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D043324:beta-Fructofuranosidase; D001616:beta-Galactosidase / Q000235:genetics
D004271:DNA, Fungal; D006026:Glycoside Hydrolases; D001616:beta-Galactosidase; D043324:beta-Fructofuranosidase; D005947:Glucose