Zwilch, a new component of the ZW10/ROD complex required for kinetochore functions.
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.
Williams
Byron C
BC
Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853-2703, USA.
Li
ZeXiao
Z
Liu
Songtao
S
Williams
Erika V
EV
Leung
Garmay
G
Yen
Tim J
TJ
Goldberg
Michael L
ML
D000595:Amino Acid Sequence; D000818:Animals; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004330:Drosophila / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D029721:Drosophila Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017344:Genes, Insect; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D018386:Kinetochores / Q000378:metabolism*; D046911:Macromolecular Substances; D008869:Microtubule-Associated Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008938:Mitosis; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D008941:Spindle Apparatus / Q000378:metabolism
D018797:Cell Cycle Proteins; D029721:Drosophila Proteins; C489309:KNTC1 protein, human; D046911:Macromolecular Substances; D008869:Microtubule-Associated Proteins; C064993:ROD protein, Drosophila; C474841:Zwilch protein, Drosophila; C076055:Zw10 protein, Drosophila