Characterization of SKM1, a Saccharomyces cerevisiae gene encoding a novel Ste20/PAK-like protein kinase

Mol Microbiol. 1997 Feb;23(3):431-44. doi: 10.1046/j.1365-2958.1997.d01-1870.x.

Abstract

Ste20/PAK serine/threonine protein kinases have been suggested as playing essential roles in cell signalling and morphogenesis as potential targets of Cdc42 and Rac GTPases. We have isolated and characterized the Saccharomyces cerevisiae SKM1 gene, which codes for a novel member of this family of protein kinases. The amino acid sequence analysis of Skm1p revealed the presence of a PH domain and a putative p21-binding domain near its amino terminus, suggesting its involvement in cellular signalling or cytoskeletal functions. However, deletion of SKM1 produced no detectable phenotype under standard laboratory conditions. Moreover, disruption of each of the two other S. cerevisiae Ste20/PAK-like kinase-encoding genes, STE20 and CLA4, in skm1 backgrounds, showed that Skm1p is not redundant with Ste20p or Cla4p. Interestingly, overexpression of SKM1 led to morphological alterations, indicating a possible role for this protein in morphogenetic control. Furthermore, overproduction of Skm1p lacking its N-terminus caused growth arrest. This effect was also seen when similarly truncated versions of Ste20p or Cla4p were overexpressed. We further observed that overproduction of this C-terminal fragment of Skm1p complements the mating defect of a ste20 mutant strain. These results suggest that the N-terminal domains of S. cerevisiae Ste20/ PAK-like protein kinases share a negative regulatory function and play a role in substrate specificity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Fungal Proteins / genetics*
  • Fungal Proteins / physiology
  • Gene Expression Regulation, Fungal
  • Genes, Fungal* / genetics
  • Genes, Fungal* / physiology
  • Genomic Library
  • Molecular Sequence Data
  • Mutagenesis
  • Peptide Fragments / genetics
  • Peptide Fragments / physiology
  • Protein Structure, Tertiary
  • Protein-Serine-Threonine Kinases / genetics*
  • Protein-Serine-Threonine Kinases / metabolism
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / physiology
  • Saccharomyces cerevisiae Proteins*
  • Sequence Analysis

Substances

  • Fungal Proteins
  • Peptide Fragments
  • Saccharomyces cerevisiae Proteins
  • SKM1 protein, S cerevisiae
  • Protein-Serine-Threonine Kinases

Associated data

  • GENBANK/X69322