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Vol. 16, No. 2, pp. 183-197, January 15, 2002
1 Verna and Marrs McLean Department of Biochemistry and
Molecular Biology, 2 Department of Cell Biology,
3 Department of Molecular and Human Genetics,
4 Howard Hughes Medical Institute, Baylor College of Medicine,
Houston, Texas 77030, USA
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 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.
[Key Words: Ask1; spindle; kinetochore; chromosome segregation]
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