A protein kinase substrate identified by the two-hybrid system

Science. 1992 Jul 31;257(5070):680-2. doi: 10.1126/science.1496382.

Abstract

A genetic method, the two-hybrid system, was used to identify four genes encoding proteins that interact with the SNF1 protein kinase from yeast. One of the genes, SIP1, was independently isolated as a multicopy suppressor of defects caused by reduced SNF1 kinase activity, and genetic evidence supports its function in the SNF1 pathway. The SIP1 protein co-immunoprecipitated with SNF1 and was phosphorylated in vitro. Thus, the two-hybrid system, which is applicable to any cloned gene, can be used to detect physical interactions between protein kinases and functionally related substrate proteins.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • AMP-Activated Protein Kinases
  • Amino Acid Sequence
  • Base Sequence
  • DNA-Binding Proteins / metabolism
  • Fungal Proteins / genetics*
  • Fungal Proteins / metabolism*
  • Genes, Fungal
  • Molecular Sequence Data
  • Mutagenesis
  • Phosphorylation
  • Plasmids
  • Protein Kinases / metabolism*
  • Protein Serine-Threonine Kinases*
  • Recombinant Fusion Proteins / metabolism*
  • Restriction Mapping
  • Saccharomyces cerevisiae / enzymology*
  • Saccharomyces cerevisiae Proteins*
  • Substrate Specificity
  • Transcription Factors*

Substances

  • DNA-Binding Proteins
  • Fungal Proteins
  • GAL4 protein, S cerevisiae
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors
  • Protein Kinases
  • SNF1-related protein kinases
  • Protein Serine-Threonine Kinases
  • SIP1 protein, S cerevisiae
  • AMP-Activated Protein Kinases