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Vol. 15, Issue 3, 1146-1159, March 2004
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* Departments of Anatomy and Cell Biology, Indiana University Medical Sciences Program, Bloomington, Indiana 47405;
Departments of Biochemistry and Molecular Biology, Indiana University Medical Sciences Program, Bloomington, Indiana 47405; and
Laboratory of Cell Regulation, Division of Molecular Medicine, Wadsworth Center, Albany, New York 12201-0509
Submitted August 11, 2003;
Revised November 20, 2003;
Accepted November 21, 2003
Monitoring Editor: Lawrence Goldstein
The complex behavior of chromosomes during mitosis is accomplished by precise binding and highly regulated polymerization dynamics of kinetochore microtubules. Previous studies have implicated Kin Is, unique kinesins that depolymerize microtubules, in regulating chromosome positioning. We have characterized the immunofluorescence localization of centromere-bound MCAK and found that MCAK localized to inner kinetochores during prophase but was predominantly centromeric by metaphase. Interestingly, MCAK accumulated at leading kinetochores during congression but not during segregation. We tested the consequences of MCAK disruption by injecting a centromere dominant-negative protein into prophase cells. Depletion of centromeric MCAK led to reduced centromere stretch, delayed chromosome congression, alignment defects, and severe missegregation of chromosomes. Rates of chromosome movement were unchanged, suggesting that the primary role of MCAK is not to move chromosomes. Furthermore, we found that disruption of MCAK leads to multiple kinetochore–microtubule attachment defects, including merotelic, syntelic, and combined merotelic-syntelic attachments. These findings reveal an essential role for Kin Is in prevention and/or correction of improper kinetochore–microtubule attachments.
Online version of this article contains supplementary figures and video material. Online version is available at www.molbiolcell.org.
Corresponding author. E-mail address: cwalczak{at}indiana.edu.
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