title: A cell cycle phosphoproteome of the yeast centrosome.
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.
authors.affiliation: Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, CO 80309, USA.
authors.forename: Jamie M
authors.initials: JM
authors.lastname: Keck
authors.forename: Michele H
authors.initials: MH
authors.lastname: Jones
authors.forename: Catherine C L
authors.initials: CC
authors.lastname: Wong
authors.forename: Jonathan
authors.initials: J
authors.lastname: Binkley
authors.forename: Daici
authors.initials: D
authors.lastname: Chen
authors.forename: Sue L
authors.initials: SL
authors.lastname: Jaspersen
authors.forename: Eric P
authors.initials: EP
authors.lastname: Holinger
authors.forename: Tao
authors.initials: T
authors.lastname: Xu
authors.forename: Mario
authors.initials: M
authors.lastname: Niepel
authors.forename: Michael P
authors.initials: MP
authors.lastname: Rout
authors.forename: Jackie
authors.initials: J
authors.lastname: Vogel
authors.forename: Arend
authors.initials: A
authors.lastname: Sidow
authors.forename: John R
authors.initials: JR
authors.lastname: Yates
authors.forename: Mark
authors.initials: M
authors.lastname: Winey
mesh_terms: D001665:Binding Sites; D016203:CDC2 Protein Kinase / Q000378:metabolism; D002453:Cell Cycle; D018385:Centrosome / Q000378:metabolism* / Q000648:ultrastructure; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism; D005658:Fungi / Q000378:metabolism; D016193:G1 Phase; D008938:Mitosis; D009154:Mutation; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008941:Spindle Apparatus / Q000378:metabolism / Q000648:ultrastructure; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism
chemical_list: D003598:Cytoskeletal Proteins; D005656:Fungal Proteins; D010750:Phosphoproteins; D020543:Proteome; C432692:SPC42 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C100764:TUB4 protein, S cerevisiae; D014404:Tubulin; D016203:CDC2 Protein Kinase