Yeast Pak1 kinase associates with and activates Snf1.

  • Nath N
  • McCartney R
  • Schmidt M
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Abstract

Members of the Snf1/AMP-activated protein kinase family are activated under conditions of nutrient stress by a distinct upstream kinase. Here we present evidence that the yeast Pak1 kinase functions as a Snf1-activating kinase. Pak1 associates with the Snf1 kinase in vivo, and the association is greatly enhanced under glucose-limiting conditions when Snf1 is active. Snf1 kinase complexes isolated from pak1Delta mutant strains show reduced specific activity in vitro, and affinity-purified Pak1 kinase is able to activate the Snf1-dependent phosphorylation of Mig1 in vitro. Purified Pak1 kinase promotes the phosphorylation of the Snf1 polypeptide on threonine 210 within the activation loop in vitro, and an increased dosage of the PAK1 gene causes increased Snf1 threonine 210 phosphorylation in vivo. Deletion of the PAK1 gene does not produce a Snf phenotype, suggesting that one or more additional protein kinases is able to activate Snf1 in vivo. However, deletion of the PAK1 gene suppresses many of the phenotypes associated with the deletion of the REG1 gene, providing genetic evidence that Pak1 activates Snf1 in vivo. The closest mammalian homologue of yeast Pak1 kinase, calcium-calmodulin-dependent protein kinase kinase beta, may play a similar role in mammalian nutrient stress signaling.

Author-supplied keywords

  • Enzyme Activation
  • Glucose
  • Glucose: metabolism
  • Macromolecular Substances
  • Phosphoprotein Phosphatases
  • Phosphoprotein Phosphatases: metabolism
  • Phosphorylation
  • Protein Phosphatase 1
  • Protein Subunits
  • Protein Subunits: metabolism
  • Protein-Serine-Threonine Kinases
  • Protein-Serine-Threonine Kinases: genetics
  • Protein-Serine-Threonine Kinases: metabolism
  • Recombinant Fusion Proteins
  • Recombinant Fusion Proteins: metabolism
  • Saccharomyces cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Saccharomyces cerevisiae Proteins: genetics
  • Saccharomyces cerevisiae Proteins: metabolism
  • Saccharomyces cerevisiae: genetics
  • Saccharomyces cerevisiae: metabolism
  • Threonine
  • Threonine: metabolism
  • p21-Activated Kinases

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Authors

  • Nandita Nath

  • Rhonda R McCartney

  • Martin C Schmidt

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