The mitotic spindle is required for loading of the DASH complex onto the kinetochore

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Abstract
A role for the mitotic spindle in the maturation of the kinetochore has not been defined previously. Here we describe the isolation of a novel and conserved essential gene, ASK1, fromSaccharomyces cerevisiae involved in this process. ask1mutants display either G2/M arrest or segregation of DNA masses without the separation of sister chromatids, resulting in massive nondisjunction and broken spindles. Ask1 localizes along mitotic spindles and to kinetochores, and cross-links to centromeric DNA. Microtubules are required for Ask1 binding to kinetochores, and are partially required to maintain its association. We found Ask1 is part of a multisubunit complex, DASH, that contains ∼10 components, including several proteins essential for mitosis including Dam1, Duo1, Spc34, Spc19, and Hsk1. The Ipl1 kinase controls the phosphorylation of Dam1 in the DASH complex and may regulate its function. We propose that DASH is a microtubule-binding complex that is transferred to the kinetochore prior to mitosis, thereby defining a new step in kinetochore maturation.