Journal
MOLECULAR BIOLOGY OF THE CELL
Volume 15, Issue 4, Pages 1580-1590Publisher
AMER SOC CELL BIOLOGY
DOI: 10.1091/mbc.E03-07-0544
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- NIGMS NIH HHS [GM58025, R01 GM058025] Funding Source: Medline
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The XMAP215/Dis1 MAP family is thought to regulate microtubule plus-end assembly in part by antagonizing the catastrophe-promoting function of kin I kinesins, yet XMAP215/Dis1 proteins localize to centrosomes. We probed the mitotic function of TOGp (human homolog of XMAP215/Dis1) using siRNA. Cells lacking TOGp assembled multipolar spindles, confirming results of Gergely et al. (2003. Genes Dev. 17, 336-341). Eg5 motor activity was necessary to maintain the multipolar morphology. Depletion of TOGp decreased microtubule length and density in the spindle by similar to20%. Depletion of MCAK, a kin I kinesin, increased NIT lengths and density by similar to20%, but did not disrupt spindle morphology. Mitotic cells lacking both TOGp and MCAK formed bipolar and monopolar spindles, indicating that TOGp and MCAK contribute to spindle bipolarity, without major effects on MT stability. TOGp localized to centrosomes in the absence of MTs and depletion of TOGp resulted in centrosome fragmentation. TOGp depletion also disrupted NIT minus-end focus at the spindle poles, detected by localizations of NuMA and the p150 component of dynactin. The major functions of TOGp during mitosis are to focus MT minus ends at spindle poles, maintain centrosorne integrity, and contribute to spindle bipolarity.
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