Depletion of centromeric MCAK leads to chromosome congression and segregation defects due to improper kinetochore attachments.
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.
Kline-Smith
Susan L
SL
Departments of Anatomy and Cell Biology, Indiana University Medical Sciences Program, Bloomington, Indiana 47405, USA.
Khodjakov
Alexey
A
Hergert
Polla
P
Walczak
Claire E
CE
D000818:Animals; D002453:Cell Cycle / Q000502:physiology; D002478:Cells, Cultured; D020090:Chromosome Segregation / Q000502:physiology*; D002875:Chromosomes / Q000378:metabolism*; D006801:Humans; D016547:Kinesins / Q000378:metabolism*; D018386:Kinetochores / Q000378:metabolism*; D008854:Microscopy, Electron; D008870:Microtubules / Q000378:metabolism; D008941:Spindle Apparatus / Q000378:metabolism; D014981:Xenopus / Q000378:metabolism
C091489:KIF2C protein, human; D016547:Kinesins