title: Subunit organization in the Dam1 kinetochore complex and its ring around microtubules.
abstract: All eukaryotic cells must segregate their chromosomes equally between two daughter cells at each division. This process needs to be robust, as errors in the form of loss or gain of genetic material have catastrophic effects on viability. Chromosomes are captured, aligned, and segregated to daughter cells via interaction with spindle microtubules mediated by the kinetochore. In Saccharomyces cerevisiae one microtubule attaches to each kinetochore, requiring extreme processivity from this single connection. The yeast Dam1 complex, an essential component of the outer kinetochore, forms rings around microtubules and in vitro recapitulates much of the functionality of a kinetochore-microtubule attachment. To understand the mechanism of the Dam1 complex at the kinetochore, we must know how it binds to microtubules, how it assembles into rings, and how assembly is regulated. We used electron microscopy to map several subunits within the structure of the Dam1 complex and identify the organization of Dam1 complexes within the ring. Of importance, new data strongly support a more passive role for the microtubule in Dam1 ring formation. Integrating this information with previously published data, we generated a structural model for the Dam1 complex assembly that advances our understanding of its function and will direct future experiments.
authors.affiliation: Biophysics Graduate Group, University of California, Berkeley, Berkeley, CA 94720, USA.
authors.forename: Vincent H
authors.initials: VH
authors.lastname: Ramey
authors.forename: Amanda
authors.initials: A
authors.lastname: Wong
authors.forename: Jie
authors.initials: J
authors.lastname: Fang
authors.forename: Stuart
authors.initials: S
authors.lastname: Howes
authors.forename: Georjana
authors.initials: G
authors.lastname: Barnes
authors.forename: Eva
authors.initials: E
authors.lastname: Nogales
mesh_terms: D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000378:metabolism*; D002455:Cell Division; D020090:Chromosome Segregation; D002875:Chromosomes / Q000378:metabolism; D018386:Kinetochores / Q000378:metabolism* / Q000648:ultrastructure; D008869:Microtubule-Associated Proteins / Q000737:chemistry* / Q000378:metabolism*; D008870:Microtubules / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D012441:Saccharomyces cerevisiae / Q000378:metabolism / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism*; D008941:Spindle Apparatus / Q000378:metabolism*
chemical_list: D018797:Cell Cycle Proteins; C116003:DAM1 protein, S cerevisiae; D008869:Microtubule-Associated Proteins; D029701:Saccharomyces cerevisiae Proteins