ADH2 expression is repressed by REG1 independently of mutations that alter the phosphorylation of the yeast transcription factor ADR1.
In Saccharomyces cerevisiae, expression of the ADH2 gene is undetectable during growth on glucose. The transcription factor ADR1 is required to fully activate expression when glucose becomes depleted. Partial activation during growth on glucose occurred in cells carrying a constitutive allele of ADR1 in which the phosphorylatable serine of a cyclic AMP (cAMP)-dependent protein kinase phosphorylation site had been changed to alanine. When glucose was removed from the growth medium, a substantial increase in the level of this constitutive expression was observed for both the ADH2 gene and a reporter construct containing the ADR1 binding site. This suggests that glucose can block ADR1-mediated activation independently of cAMP-dependent phosphorylation at serine 230. REG1/HEX2/SRN1 was identified as a potential serine 230-independent repressor of ADH2 expression. Yeast strains carrying a deletion of the REG1 gene, reg1-1966, showed a large increase in ADR1-dependent expression of ADH2 during growth on glucose. A smaller increase in ADR1-independent expression was also observed. Additionally, an increase in the level of ADR1 expression and posttranslational modification of the ADR1 protein were observed. When the reg1-1966 allele was combined with various ADR1 constitutive alleles, the level of ADH2 expression was synergistically elevated. This indicates that REG1 can act independently of phosphorylation at serine 230. Our results suggest that glucose repression in the presence of ADR1 constitutive alleles occurs primarily through a REG1-dependent pathway which appears to affect ADH2 transcription at multiple levels.
Dombek
K M
KM
Department of Biochemistry, University of Washington, Seattle 98195.
Camier
S
S
Young
E T
ET
D000426:Alcohol Dehydrogenase / Q000235:genetics*; D000483:Alleles; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D003001:Cloning, Molecular; D004271:DNA, Fungal; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005656:Fungal Proteins / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D005947:Glucose / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D009154:Mutation; D010766:Phosphorylation; D012097:Repressor Proteins / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
C074682:ADR1 protein, S cerevisiae; D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D000426:Alcohol Dehydrogenase; D005947:Glucose