Protein kinase Byr2 is a target of Ras1 in the fission yeast Schizosaccharomyces pombe

J Biol Chem. 1995 Feb 3;270(5):1979-82. doi: 10.1074/jbc.270.5.1979.

Abstract

Conservation of the structure and function of Ras proteins has been observed in a variety of eukaryotic organisms. However, the nature of their downstream effectors appears to be quite divergent; adenylyl cyclase and a protein kinase Raf-1, which do not share any structural homology with each other, are effectors of Ras in the budding yeast and in higher organisms, respectively. We show here that a protein kinase Byr2, which has been known to act downstream of Ras1 in a mating pheromone signal transduction system of Schizosaccharomyces pombe, binds directly to Ras proteins in a GTP-dependent manner. The region of Byr2 responsible for the Ras binding was mapped by a gene deletion analysis to its N-terminal segment of 206 amino acid residues, which does not possess any significant homology with the other effectors of Ras. The affinity of the Byr2 N terminus for Saccharomyces cerevisiae Ras2 was determined by measuring its activity to competitively inhibit Ras-dependent adenylyl cyclase activity and found to be comparable with those of yeast adenylyl cyclase and human Raf-1, with a dissociation constant (Kd) of about 1 nM. Furthermore, Byr2 inhibited a Ras GTPase-activating activity of Ira2, a S. cerevisiae homologue of neurofibromin. These results indicate that Byr2 is an immediate downstream target of Ras1 in S. pombe.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenylyl Cyclases / metabolism
  • Enzyme Activation
  • Fungal Proteins / metabolism
  • Fungal Proteins / physiology*
  • GTPase-Activating Proteins
  • Guanosine Triphosphate / metabolism
  • Protein Kinases / physiology*
  • Protein Serine-Threonine Kinases / metabolism
  • Proteins / metabolism
  • Proto-Oncogene Proteins / metabolism
  • Proto-Oncogene Proteins c-raf
  • Saccharomyces cerevisiae Proteins*
  • Schizosaccharomyces / enzymology*
  • Schizosaccharomyces pombe Proteins*
  • ras GTPase-Activating Proteins
  • ras Proteins*

Substances

  • Fungal Proteins
  • GTPase-Activating Proteins
  • IRA2 protein, S cerevisiae
  • Proteins
  • Proto-Oncogene Proteins
  • Saccharomyces cerevisiae Proteins
  • Schizosaccharomyces pombe Proteins
  • ras GTPase-Activating Proteins
  • Guanosine Triphosphate
  • Protein Kinases
  • Protein Serine-Threonine Kinases
  • Proto-Oncogene Proteins c-raf
  • RAS1 protein, S cerevisiae
  • RAS2 protein, S cerevisiae
  • Ras1 protein, S pombe
  • ras Proteins
  • Adenylyl Cyclases