Regulation of yeast actin cytoskeleton-regulatory complex Pan1p/Sla1p/End3p by serine/threonine kinase Prk1p

Mol Biol Cell. 2001 Dec;12(12):3759-72. doi: 10.1091/mbc.12.12.3759.

Abstract

The serine/threonine kinase Prk1p is known to be involved in the regulation of the actin cytoskeleton organization in budding yeast. One possible function of Prk1p is the negative regulation of Pan1p, an actin patch regulatory protein that forms a complex in vivo with at least two other proteins, Sla1p and End3p. In this report, we identified Sla1p as another substrate for Prk1p. The phosphorylation of Sla1p by Prk1p was established in vitro with the use of immunoprecipitated Prk1p and in vivo with the use of PRK1 overexpression, and was further supported by the finding that immunoprecipitated Sla1p contained PRK1- and ARK1-dependent kinase activities. Stable complex formation between Prk1p and Sla1p/Pan1p in vivo could be observed once the phosphorylation reaction was blocked by mutation in the catalytic site of Prk1p. Elevation of Prk1p activities in wild-type cells resulted in a number of deficiencies, including those in colocalization of Pan1p and Sla1p, endocytosis, and cell wall morphogenesis, likely attributable to a disintegration of the Pan1p/Sla1p/End3p complex. These results lend a strong support to the model that the phosphorylation of the Pan1p/Sla1p/End3p complex by Prk1p is one of the important mechanisms by which the organization and functions of the actin cytoskeleton are regulated.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism*
  • Amino Acid Sequence
  • Blotting, Western
  • Carrier Proteins / metabolism*
  • Cell Wall / metabolism
  • Cell Wall / pathology
  • Chromatography, Gel
  • Cytoskeletal Proteins*
  • Cytoskeleton / metabolism*
  • Fungal Proteins / metabolism*
  • Macromolecular Substances
  • Microfilament Proteins
  • Microscopy, Electron
  • Phosphorylation
  • Protein Binding
  • Protein Kinase C
  • Protein-Serine-Threonine Kinases / genetics
  • Protein-Serine-Threonine Kinases / metabolism*
  • Receptor Protein-Tyrosine Kinases / genetics
  • Receptor Protein-Tyrosine Kinases / metabolism*
  • Saccharomyces cerevisiae / cytology*
  • Saccharomyces cerevisiae / enzymology*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / ultrastructure
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism
  • Two-Hybrid System Techniques

Substances

  • Actins
  • Carrier Proteins
  • Cytoskeletal Proteins
  • END3 protein, S cerevisiae
  • Fungal Proteins
  • Macromolecular Substances
  • Microfilament Proteins
  • PAN1 protein, S cerevisiae
  • SLA1 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • protein kinase N
  • Receptor Protein-Tyrosine Kinases
  • Protein-Serine-Threonine Kinases
  • Protein Kinase C