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Published online February 11, 2008
doi:10.1083/jcb.200708021
The Journal of Cell Biology, Vol. 180, No. 3, 507-520
The Rockefeller University Press, 0021-9525 $30.00
© 2008 Famulski et al.
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Article

 Stable hZW10 kinetochore residency, mediated by hZwint-1 interaction, is essential for the mitotic checkpoint

Jakub K. Famulski1, Larissa Vos1, Xuejun Sun2, and Gordon Chan1,2

1 Department of Oncology, University of Alberta, Edmonton, Alberta, Canada T6G 1Z2
2 Experimental Oncology, Cross Cancer Institute, Edmonton, Alberta, Canada T6G 1Z2

Correspondence to Gordon Chan: gordonch{at}cancerboard.ab.ca

The mitotic checkpoint is an essential surveillance mechanism that ensures high fidelity chromosome segregation during mitosis. Mitotic checkpoint function depends on numerous kinetochore proteins, including ZW10, ROD, and Zwilch (the ROD–ZW10–Zwilch complex). Through an extensive mutagenesis screen of hZW10, we have mapped the kinetochore localization domain of hZW10 as well as the hZwint-1 interaction domain. We find that hZwint-1–noninteracting mutants still localize to kinetochores. In addition, using fluorescence recovery after photobleaching, we have found that hZW10 residency at metaphase kinetochores is brief (half-time of 13 s). However, during prometaphase or at unattached kinetochores, enhanced green fluorescent protein–hZW10 becomes a stable component of the kinetochore. Moreover, we find that stable hZW10 kinetochore residency at prometaphase kinetochores is dependent on its interaction with hZwint-1, and is essential for mitotic checkpoint arrest.

Abbreviations used in this paper: ACA, anticentromere antibody; MT, microtubule; PEI, polyethyleneimine; RZZ, ROD–ZW10–Zwilch.


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