title: Phosphorylation of centromeric histone H3 variant regulates chromosome segregation in Saccharomyces cerevisiae.
abstract: The centromeric histone H3 variant (CenH3) is essential for chromosome segregation in eukaryotes. We identify posttranslational modifications of Saccharomyces cerevisiae CenH3, Cse4. Functional characterization of cse4 phosphorylation mutants shows growth and chromosome segregation defects when combined with kinetochore mutants okp1 and ame1. Using a phosphoserine-specific antibody, we show that the association of phosphorylated Cse4 with centromeres increases in response to defective microtubule attachment or reduced cohesion. We determine that evolutionarily conserved Ipl1/Aurora B contributes to phosphorylation of Cse4, as levels of phosphorylated Cse4 are reduced at centromeres in ipl1 strains in vivo, and in vitro assays show phosphorylation of Cse4 by Ipl1. Consistent with these results, we observe that a phosphomimetic cse4-4SD mutant suppresses the temperature-sensitive growth of ipl1-2 and Ipl1 substrate mutants dam1 spc34 and ndc80, which are defective for chromosome biorientation. Furthermore, cell biology approaches using a green fluorescent protein-labeled chromosome show that cse4-4SD suppresses chromosome segregation defects in dam1 spc34 strains. On the basis of these results, we propose that phosphorylation of Cse4 destabilizes defective kinetochores to promote biorientation and ensure faithful chromosome segregation. Taken together, our results provide a detailed analysis, in vivo and in vitro, of Cse4 phosphorylation and its role in promoting faithful chromosome segregation.
authors: Genetics Branch, Center for Cancer Research, National Institutes of Health, Bethesda, MD 20892, USA.
authors: Lars
authors: L
authors: Boeckmann
authors: Yoshimitsu
authors: Y
authors: Takahashi
authors: Wei-Chun
authors: WC
authors: Au
authors: Prashant K
authors: PK
authors: Mishra
authors: John S
authors: JS
authors: Choy
authors: Anthony R
authors: AR
authors: Dawson
authors: May Y
authors: MY
authors: Szeto
authors: Timothy J
authors: TJ
authors: Waybright
authors: Christopher
authors: C
authors: Heger
authors: Christopher
authors: C
authors: McAndrew
authors: Paul K
authors: PK
authors: Goldsmith
authors: Timothy D
authors: TD
authors: Veenstra
authors: Richard E
authors: RE
authors: Baker
authors: Munira A
authors: MA
authors: Basrai
mesh_terms: D000595:Amino Acid Sequence; D064107:Aurora Kinase B / Q000235:genetics / Q000378:metabolism; D064127:Aurora Kinases / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites / Q000235:genetics; D015153:Blotting, Western; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D002503:Centromere / Q000235:genetics / Q000378:metabolism*; D002853:Chromatography, Liquid; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism*; D020090:Chromosome Segregation; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D018386:Kinetochores / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D053719:Tandem Mass Spectrometry
chemical_list: C509718:Ame1 protein, S cerevisiae; C092746:CSE4 protein, S cerevisiae; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; C116003:DAM1 protein, S cerevisiae; D004268:DNA-Binding Proteins; D008869:Microtubule-Associated Proteins; C112472:NDC80 protein, S cerevisiae; D009687:Nuclear Proteins; C509719:Okp1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C487376:Spc34 protein, S cerevisiae; D064107:Aurora Kinase B; D064127:Aurora Kinases; C576961:IPL1 protein, S cerevisiae
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