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A cell cycle phosphoproteome of the yeast centrosome.

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.

Pubmed ID: 21700874

Authors

  • Keck JM
  • Jones MH
  • Wong CC
  • Binkley J
  • Chen D
  • Jaspersen SL
  • Holinger EP
  • Xu T
  • Niepel M
  • Rout MP
  • Vogel J
  • Sidow A
  • Yates JR
  • Winey M

Journal

Science (New York, N.Y.)

Publication Data

June 24, 2011

Associated Grants

  • Agency: NIGMS NIH HHS, Id: F32 GM086038
  • Agency: NIGMS NIH HHS, Id: GM51312
  • Agency: Canadian Institutes of Health Research, Id: MOP-64404
  • Agency: NCRR NIH HHS, Id: P41 RR011823
  • Agency: NIGMS NIH HHS, Id: R01 GM051312
  • Agency: NIGMS NIH HHS, Id: R01 GM051312-16
  • Agency: NIGMS NIH HHS, Id: R01 GM051312-16S1
  • Agency: NIGMS NIH HHS, Id: R01 GM062427
  • Agency: NIGMS NIH HHS, Id: R01 GM062427
  • Agency: NHGRI NIH HHS, Id: R01 HG003039
  • Agency: NIGMS NIH HHS, Id: T32 GM008759
  • Agency: NCRR NIH HHS, Id: U54 RR022220

Mesh Terms

  • Binding Sites
  • CDC2 Protein Kinase
  • Cell Cycle
  • Centrosome
  • Cytoskeletal Proteins
  • Fungal Proteins
  • Fungi
  • G1 Phase
  • Mitosis
  • Mutation
  • Phosphoproteins
  • Phosphorylation
  • Protein Processing, Post-Translational
  • Proteome
  • Saccharomyces cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Spindle Apparatus
  • Tubulin