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Vol. 14, Issue 4, 1379-1391, April 2003

and
*Department of Molecular Biology and Genetics, Cornell
University, Ithaca, New York 14853-2703; and The Zeste-White 10 (ZW10) and Rough Deal (ROD) proteins are part of
a complex necessary for accurate chromosome segregation. This complex
recruits cytoplasmic dynein to the kinetochore and participates in the spindle checkpoint. We used immunoaffinity chromatography and mass spectroscopy to identify the
Drosophila proteins in this complex. We found that the
complex contains an additional protein we name Zwilch. Zwilch localizes
to kinetochores and kinetochore microtubules in
a manner identical to ZW10 and ROD. We have also isolated a
zwilch mutant, which exhibits the same mitotic
phenotypes associated with zw10 and rod
mutations: lagging chromosomes at anaphase and precocious sister
chromatid separation upon activation of the spindle checkpoint.
Zwilch's role within the context of this complex is evolutionarily
conserved. The human Zwilch protein (hZwilch) coimmunoprecipitates with
hZW10 and hROD from HeLa cell extracts and localizes to the
kinetochores at prometaphase. Finally, we discuss
immunoaffinity chromatography results that suggest the existence of a
weak interaction between the ZW10/ROD/Zwilch complex and the
kinesin-like kinetochore component CENP-meta.
Fox
Chase Cancer Center, Philadelphia, Pennsylvania 19111
Corresponding author. E-mail address:
mlg11{at}cornell.edu.
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