Histone H3 Ser10 phosphorylation-independent function of Snf1 and Reg1 proteins rescues a gcn5- mutant in HIS3 expression

Mol Cell Biol. 2005 Dec;25(23):10566-79. doi: 10.1128/MCB.25.23.10566-10579.2005.

Abstract

Gcn5 protein is a prototypical histone acetyltransferase that controls transcription of multiple yeast genes. To identify molecular functions that act downstream of or in parallel with Gcn5 protein, we screened for suppressors that rescue the transcriptional defects of HIS3 caused by a catalytically inactive mutant Gcn5, the E173H mutant. One bypass of Gcn5 requirement gene (BGR) suppressor was mapped to the REG1 locus that encodes a semidominant mutant truncated after amino acid 740. Reg1(1-740) protein does not rescue the complete knockout of GCN5, nor does it suppress other gcn5- defects, including the inability to utilize nonglucose carbon sources. Reg1(1-740) enhances HIS3 transcription while HIS3 promoter remains hypoacetylated, indicating that a noncatalytic function of Gcn5 is targeted by this suppressor protein. Reg1 protein is a major regulator of Snf1 kinase that phosphorylates Ser10 of histone H3. However, whereas Snf1 protein is important for HIS3 expression, replacing Ser10 of H3 with alanine or glutamate neither attenuates nor augments the BGR phenotypes. Overproduction of Snf1 protein also preferentially rescues the E173H allele. Biochemically, both Snf1 and Reg1(1-740) proteins copurify with Gcn5 protein. Snf1 can phosphorylate recombinant Gcn5 in vitro. Together, these data suggest that Reg1 and Snf1 proteins function in an H3 phosphorylation-independent pathway that also involves a noncatalytic role played by Gcn5 protein.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Alleles
  • Amino Acid Sequence
  • Base Sequence
  • Gene Expression Regulation, Fungal
  • Glutamic Acid / genetics
  • Glutamic Acid / metabolism
  • Histone Acetyltransferases / deficiency*
  • Histone Acetyltransferases / genetics
  • Histone Acetyltransferases / metabolism*
  • Histones / chemistry
  • Histones / metabolism*
  • Hydro-Lyases / genetics*
  • Hydro-Lyases / metabolism
  • Molecular Sequence Data
  • Mutation / genetics
  • Phenotype
  • Phosphoprotein Phosphatases / chemistry
  • Phosphoprotein Phosphatases / genetics
  • Phosphoprotein Phosphatases / metabolism*
  • Phosphoserine / metabolism*
  • Protein Binding
  • Protein Phosphatase 1
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / genetics*
  • Saccharomyces cerevisiae Proteins / metabolism
  • Suppression, Genetic / genetics

Substances

  • Histones
  • Saccharomyces cerevisiae Proteins
  • Phosphoserine
  • Glutamic Acid
  • GCN5 protein, S cerevisiae
  • Histone Acetyltransferases
  • SNF1-related protein kinases
  • Protein Serine-Threonine Kinases
  • Phosphoprotein Phosphatases
  • Protein Phosphatase 1
  • REG1 protein, S cerevisiae
  • Hydro-Lyases
  • imidazoleglycerolphosphate dehydratase