title: Pat1 protects centromere-specific histone H3 variant Cse4 from Psh1-mediated ubiquitination.
abstract: Evolutionarily conserved histone H3 variant Cse4 and its homologues are essential components of specialized centromere (CEN)-specific nucleosomes and serve as an epigenetic mark for CEN identity and propagation. Cse4 is a critical determinant for the structure and function of the kinetochore and is required to ensure faithful chromosome segregation. The kinetochore protein Pat1 regulates the levels and spatial distribution of Cse4 at centromeres. Deletion of PAT1 results in altered structure of CEN chromatin and chromosome segregation errors. In this study, we show that Pat1 protects CEN-associated Cse4 from ubiquitination in order to maintain proper structure and function of the kinetochore in budding yeast. PAT1-deletion strains exhibit increased ubiquitination of Cse4 and faster turnover of Cse4 at kinetochores. Psh1, a Cse4-specific E3-ubiquitin ligase, interacts with Pat1 in vivo and contributes to the increased ubiquitination of Cse4 in pat1∆ strains. Consistent with a role of Psh1 in ubiquitination of Cse4, transient induction of PSH1 in a wild-type strain resulted in phenotypes similar to a pat1∆ strain, including a reduction in CEN-associated Cse4, increased Cse4 ubiquitination, defects in spatial distribution of Cse4 at kinetochores, and altered structure of CEN chromatin. Pat1 interacts with Scm3 and is required for its maintenance at kinetochores. In conclusion, our studies provide novel insights into mechanisms by which Pat1 affects the structure of CEN chromatin and protects Cse4 from Psh1-mediated ubiquitination for faithful chromosome segregation.
authors.affiliation: Genetics Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
authors.forename: Prashant K
authors.initials: PK
authors.lastname: Mishra
authors.affiliation: Department of Biology, University of North Carolina, Chapel Hill, NC 27599.
authors.forename: Jiasheng
authors.initials: J
authors.lastname: Guo
authors.affiliation: Genetics Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
authors.forename: Lauren E
authors.initials: LE
authors.lastname: Dittman
authors.affiliation: Department of Biology, University of North Carolina, Chapel Hill, NC 27599.
authors.forename: Julian
authors.initials: J
authors.lastname: Haase
authors.affiliation: Department of Biology, University of North Carolina, Chapel Hill, NC 27599.
authors.forename: Elaine
authors.initials: E
authors.lastname: Yeh
authors.affiliation: Department of Biology, University of North Carolina, Chapel Hill, NC 27599.
authors.forename: Kerry
authors.initials: K
authors.lastname: Bloom
authors.affiliation: Genetics Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892 basraim@mail.nih.gov.
authors.forename: Munira A
authors.initials: MA
authors.lastname: Basrai
mesh_terms: D002503:Centromere / Q000378:metabolism*; D002843:Chromatin / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006657:Histones / Q000378:metabolism*; D010445:Peptide Elongation Factors / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000502:physiology*; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism* / Q000502:physiology*; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*; D054875:Ubiquitination
chemical_list: C092746:CSE4 protein, S cerevisiae; D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D006657:Histones; C103266:PAT1 protein, S cerevisiae; D010445:Peptide Elongation Factors; D016601:RNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; C554184:Psh1 protein, S cerevisiae; D044767:Ubiquitin-Protein Ligases
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