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MBC in Press, published online ahead of print June 13, 2003
Mol. Biol. Cell 10.1091/mbc.E03-02-0082

A more recent version of this article appeared on September 1, 2003
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Submitted on February 13, 2003
Revised on April 25, 2003
Accepted on May 19, 2003

A Functional Relationship between NuMA and Kid is involved in both spindle organization and chromosome alignment in vertebrate cells

Aime A. Levesque1, Louisa Howard2, Michael B. Gordon1, and Duane A. Compton1*

1 Department of Biochemistry, Dartmouth Medical School, Hanover, N.H. 03755
2 Rippel Electron Microscope Facility, Dartmouth College, Hanover, N.H. 03755

* Corresponding author. E-mail address: duane.a.compton{at}dartmouth.edu.

We examined spindle morphology and chromosome alignment in vertebrate cells following simultaneous perturbation of the chromokinesin Kid and either NuMA, CENP-E, or HSET. Spindle morphology and chromosome alignment following simultaneous perturbation of Kid and either HSET or CENP-E were no different than when either HSET or CENP-E were perturbed alone. However, short, bipolar spindles with organized poles formed following perturbation of both Kid and NuMA in stark contrast to splayed spindle poles observed following perturbation of NuMA alone. Spindles were disorganized if Kid, NuMA and HSET were perturbed indicating that HSET is sufficient for spindle organization in the absence of Kid and NuMA function. In addition, chromosomes failed to align efficiently at the spindle equator following simultaneous perturbation of Kid and NuMA despite appropriate kinetochore-microtubule interactions that generated chromosome movement at normal velocities. These data indicate that a functional relationship between the chromokinesin Kid and the spindle pole organizing protein NuMA influences spindle morphology, and we propose that this occurs because NuMA forms functional linkages between kinetochore and nonkinetochore microtubules at spindle poles. In addition, these data show that both Kid and NuMA contribute to chromosome alignment in mammalian cells.




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