A cell cycle phosphoproteome of the yeast centrosome

Science. 2011 Jun 24;332(6037):1557-61. doi: 10.1126/science.1205193.

Abstract

Centrosomes organize the bipolar mitotic spindle, and centrosomal defects cause chromosome instability. Protein phosphorylation modulates centrosome function, and we provide a comprehensive map of phosphorylation on intact yeast centrosomes (18 proteins). Mass spectrometry was used to identify 297 phosphorylation sites on centrosomes from different cell cycle stages. We observed different modes of phosphoregulation via specific protein kinases, phosphorylation site clustering, and conserved phosphorylated residues. Mutating all eight cyclin-dependent kinase (Cdk)-directed sites within the core component, Spc42, resulted in lethality and reduced centrosomal assembly. Alternatively, mutation of one conserved Cdk site within γ-tubulin (Tub4-S360D) caused mitotic delay and aberrant anaphase spindle elongation. Our work establishes the extent and complexity of this prominent posttranslational modification in centrosome biology and provides specific examples of phosphorylation control in centrosome function.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • CDC2 Protein Kinase / metabolism
  • Cell Cycle*
  • Centrosome / metabolism*
  • Centrosome / ultrastructure
  • Cytoskeletal Proteins / genetics
  • Cytoskeletal Proteins / metabolism
  • Fungal Proteins / chemistry
  • Fungal Proteins / metabolism
  • Fungi / metabolism
  • G1 Phase
  • Mitosis
  • Mutation
  • Phosphoproteins / genetics
  • Phosphoproteins / metabolism
  • Phosphorylation
  • Protein Processing, Post-Translational
  • Proteome / metabolism*
  • Saccharomyces cerevisiae / cytology
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / growth & development
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Spindle Apparatus / metabolism
  • Spindle Apparatus / ultrastructure
  • Tubulin / chemistry
  • Tubulin / metabolism

Substances

  • Cytoskeletal Proteins
  • Fungal Proteins
  • Phosphoproteins
  • Proteome
  • SPC42 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • TUB4 protein, S cerevisiae
  • Tubulin
  • CDC2 Protein Kinase