Snf1/AMPK promotes SBF and MBF-dependent transcription in budding yeast

Biochim Biophys Acta. 2013 Dec;1833(12):3254-3264. doi: 10.1016/j.bbamcr.2013.09.014. Epub 2013 Sep 29.

Abstract

Snf1, the yeast AMP-activated kinase homolog, regulates the expression of several genes involved in adaptation to glucose limitation and in response to cellular stresses. We previously demonstrated that Snf1 interacts with Swi6, the regulatory subunit of SBF and MBF complexes, and activates CLB5 transcription. Here we report that, in α-factor synchronized cells in 2% glucose, the loss of the Snf1 catalytic subunit impairs the binding of SBF and MBF complexes and the subsequent recruitment of the FACT complex and RNA Polymerase II to promoters of G1-genes. By using an analog-sensitive allele of SNF1, SNF1(as)(I132G), encoding a protein whose catalytic activity is selectively inhibited in vivo by 2-naphthylmethyl pyrazolopyrimidine 1, we show that the inhibition of Snf1 catalytic activity affects the expression of G1-genes causing a delayed entrance into S phase in cells synchronized in G1 phase by α-factor treatment or by elutriation. Moreover, Snf1 is detected in immune complexes of Rpb1, the large subunit of RNA Polymerase II, and is present at both promoters and coding regions of SBF- and MBF-regulated genes 20min after α-factor release, suggesting a direct role for Snf1 in the activation of the G1-regulon transcription.

Keywords: Cell cycle; RNA Polymerase II; SBF–MBF complexes; SNF1(as)(I132G); Saccharomyces cerevisiae; Snf1/AMPK.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AMP-Activated Protein Kinases / metabolism*
  • Biocatalysis / drug effects
  • Gene Expression Regulation, Fungal / drug effects
  • Genes, Fungal / genetics
  • Glucose / pharmacology
  • Models, Biological
  • Phosphorylation / drug effects
  • Phosphothreonine / metabolism
  • Promoter Regions, Genetic / genetics
  • Protein Binding / drug effects
  • Protein Serine-Threonine Kinases / metabolism*
  • RNA Polymerase II / metabolism
  • Saccharomyces cerevisiae / drug effects
  • Saccharomyces cerevisiae / enzymology*
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / growth & development
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Transcription Factors / metabolism*
  • Transcription, Genetic* / drug effects

Substances

  • MBP1 protein, S cerevisiae
  • SBF protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors
  • Phosphothreonine
  • SNF1-related protein kinases
  • Protein Serine-Threonine Kinases
  • AMP-Activated Protein Kinases
  • RNA Polymerase II
  • Glucose