The mitotic spindle is required for loading of the DASH complex onto the kinetochore.
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, from Saccharomyces cerevisiae involved in this process. ask1 mutants display either G(2)/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 approximately 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.
Li
Yumei
Y
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas 77030, USA.
Bachant
Jeff
J
Alcasabas
Annette A
AA
Wang
Yanchang
Y
Qin
Jun
J
Elledge
Stephen J
SJ
D000595:Amino Acid Sequence; D000818:Animals; D029681:Arabidopsis Proteins; D001483:Base Sequence; D002842:Chromatids; D017931:DNA Primers; D005656:Fungal Proteins / Q000235:genetics; D016195:G2 Phase; D020043:Genes, Essential; D005800:Genes, Fungal; D018386:Kinetochores; D008938:Mitosis; D008969:Molecular Sequence Data; D009154:Mutation; D010766:Phosphorylation; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D008941:Spindle Apparatus
C120363:ASK1 protein, Arabidopsis; D029681:Arabidopsis Proteins; D017931:DNA Primers; D005656:Fungal Proteins; D010940:Plant Proteins