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MBC in Press, published online ahead of print September 21, 2005
Mol. Biol. Cell 10.1091/mbc.E05-03-0244

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Submitted on March 23, 2005
Revised on September 6, 2005
Accepted on September 8, 2005

The Chromokinesin Kid Is Required for Maintenance of Proper Metaphase Spindle Size

Noriko Tokai-Nishizumi,* Miho Ohsugi,* Emiko Suzuki,{dagger} and Tadashi Yamamoto*

*Division of Oncology, Department of Cancer Biology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan; {dagger}Gene Network Laboratory, Structural Biology Center, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan

Monitoring Editor: Kerry Bloom

The human chromokinesin Kid/kinesin-10, a plus end-directed microtubule-based motor with both microtubule- and DNA-binding domains, is required for proper chromosome alignment at the metaphase plate. Here we performed RNA interference experiments to deplete endogenous Kid from HeLa cells and confirmed defects in metaphase chromosome arm alignment in Kid-depleted cells. In addition we noted a shortening of the spindle length, resulting in a pole-to-pole distance only 80% of wild-type. The spindle microtubule-bundles with which Kid normally colocalizes became less robust. Rescue of the two Kid deficiency phenotypes--imprecise chromosome alignment at metaphase and shortened spindles--exhibited distinct requirements. Mutants lacking either the DNA-binding domain or the MT motor ATPase failed to rescue the former defect, while rescue of the shortened spindle phenotype required neither activity. Kid also exhibits microtubule bundling activity in vitro, and rescue of the shortened spindle phenotype and the bundling activity displayed similar domain requirements except that rescue required a coiled-coil domain not needed for bundling. These results suggest that distinct from its role in chromosome movement, Kid contributes to spindle morphogenesis by mediating spindle microtubules stabilization.

Address correspondence to: Tadashi Yamamoto (tyamamot{at}ims.u-tokyo.ac.jp)




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