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MBC in Press, published online ahead of print April 20, 2005
Mol. Biol. Cell 10.1091/mbc.E05-02-0167

A more recent version of this article appeared on July 1, 2005
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Submitted on February 28, 2005
Accepted on April 13, 2005

Functional Analysis of Human Microtubule-based Motor Proteins, the Kinesins and Dyneins, in Mitosis/Cytokinesis Using RNA Interference (RNAi)

Changjun Zhu,*{dagger} Jian Zhao,*{dagger}{ddagger} Marina Bibikova,{sect} Joel D. Leverson,|| Ella Bossy-Wetzel,* Jian-Bing Fan,{sect} Robert T. Abraham,* and Wei Jiang*

*The Burnham Institute, La Jolla, CA 92037; {sect}Illumina, San Diego, CA 92121; ||Abbott Laboratories, Abbott Park, IL 60064

Monitoring Editor: J. Richard McIntosh

Microtubule (MT)-based motor proteins, kinesins and dyneins, play important roles in multiple cellular processes including cell division. In this study, we describe the generation and use of an E. coli RNase III-prepared human kinesin/dynein esiRNA library to systematically analyze the functions of all human kinesin/dynein MT motor proteins. Our results indicate that at least 12 kinesins are involved in mitosis and cytokinesis. Eg5 (a member of the kinesin-5 family), Kif2A (a member of the kinesin-13 family) and KifC1 (a member of the kinesin-14 family) are crucial for spindle formation; KifC1, MCAK (a member of the kinesin-13 family), CENP-E (a member of the kinesin-7 family), Kif14 (a member of the kinesin-3 family), Kif18 (a member of the kinesin-8 family), and Kid (a member of the kinesin-10 family) are required for chromosome congression and alignment; Kif4A and Kif4B (members of the kinesin-4 family) have roles in anaphase spindle dynamics; and Kif4A, Kif4B, MKLP1 and MKLP2 (members of the kinesin-6 family) are essential for cytokinesis. Using immunofluorescence analysis, time-lapse microscopy, and rescue experiments, we investigate the roles of these 12 kinesins in detail.

{dagger}These authors contributed equally to this work.

{ddagger}Present address: Qilu Hospital, Shandong University, Jinan, Shandong, 250012 China.

Address correspondence to: Wei Jiang (wjiang{at}burnham.org)




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