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Vol. 13, Issue 8, 2718-2731, August 2002
Departments of *Anatomy and Cell Biology and
The dynamic activities of cellular microtubules (MTs) are tightly
regulated by a balance between MT-stabilizing and -destabilizing proteins. Studies in Xenopus egg extracts have shown
that the major MT destabilizer during interphase and mitosis is the
kinesin-related protein XKCM1, which depolymerizes MT ends in an
ATP-dependent manner. Herein, we examine the effects of both
overexpression and inhibition of XKCM1 on the regulation of MT dynamics
in vertebrate somatic cells. We found that XKCM1 is a MT-destabilizing
enzyme in PtK2 cells and that XKCM1 modulates cellular MT dynamics. Our results indicate that perturbation of XKCM1 levels alters the catastrophe frequency and the rescue frequency of cellular MTs. In
addition, we found that overexpression of XKCM1 or inhibition of KCM1
during mitosis leads to the formation of aberrant spindles and a
mitotic delay. The predominant spindle defects from excess XKCM1
included monoastral and monopolar spindles, as well as small prometaphase-like spindles with improper chromosomal attachments. Inhibition of KCM1 during mitosis led to prometaphase spindles with
excessively long MTs and spindles with partially separated poles and a
radial MT array. These results show that KCM1 plays a critical role in
regulating both interphase and mitotic MT dynamics in mammalian cells.
Biochemistry and Molecular Biology, Indiana University
Medical Sciences Program, Bloomington, Indiana 47405
Corresponding author. E-mail address:
cwalczak{at}indiana.edu.
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