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Published 11 April 2005. doi:10.1083/jcb.200411118
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 169, Number 1, 49-60
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Article

ZW10 links mitotic checkpoint signaling to the structural kinetochore



Geert J.P.L. Kops1, Yumi Kim1, Beth A.A. Weaver1, Yinghui Mao1, Ian McLeod2, John R. Yates, III2, Mitsuo Tagaya3, and Don W. Cleveland1

1 Ludwig Institute for Cancer Research and Department of Cellular and Molecular Medicine, University of California at San Diego, La Jolla, CA 92093
2 Department of Cell Biology, The Scripps Research Institute, La Jolla, CA 92037
3 School of Life Science, Tokyo University of Pharmacy and Life Science, Hachioji, Tokyo, Japan

Correspondence to Don W. Cleveland: dcleveland{at}ucsd.edu

The mitotic checkpoint ensures that chromosomes are divided equally between daughter cells and is a primary mechanism preventing the chromosome instability often seen in aneuploid human tumors. ZW10 and Rod play an essential role in this checkpoint. We show that in mitotic human cells ZW10 resides in a complex with Rod and Zwilch, whereas another ZW10 partner, Zwint-1, is part of a separate complex of structural kinetochore components including Mis12 and Ndc80–Hec1. Zwint-1 is critical for recruiting ZW10 to unattached kinetochores. Depletion from human cells or Xenopus egg extracts is used to demonstrate that the ZW10 complex is essential for stable binding of a Mad1–Mad2 complex to unattached kinetochores. Thus, ZW10 functions as a linker between the core structural elements of the outer kinetochore and components that catalyze generation of the mitotic checkpoint-derived "stop anaphase" inhibitor.

G.J.P.L. Kops's present address is Dept. of Experimental Oncology, UMC Utrecht, 3584 CG Utrecht, Netherlands.

Abbreviations used in this paper: ACA, anti-centromere antiserum; CENP, centromere-associated protein; CSF, cytostatic factor; EYFP, enhanced YFP; KCR, kinetochore-containing region; LAP, localization and affinity purification; siRNA, small interfering RNA; Zwint-1, ZW10-interacting protein 1.


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