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Vol. 14, Issue 9, 3541-3552, September 2003
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* Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03755;
Rippel Electron Microscope Facility, Dartmouth College, Hanover, New Hampshire 03755
Submitted February 13, 2003;
Revised April 25, 2003;
Accepted May 19, 2003
Monitoring Editor: Ted Salmon
We examined spindle morphology and chromosome alignment in vertebrate cells after simultaneous perturbation of the chromokinesin Kid and either NuMA, CENP-E, or HSET. Spindle morphology and chromosome alignment after simultaneous perturbation of Kid and either HSET or CENP-E were no different from when either HSET or CENP-E was perturbed alone. However, short bipolar spindles with organized poles formed after perturbation of both Kid and NuMA in stark contrast to splayed spindle poles observed after 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 after 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.
Online version of this article contains video material for some figures. Online version available at www.molbiolcell.org.
Corresponding author. E-mail address: duane.a.compton{at}dartmouth.edu.
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