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PMID: 12181341 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The microtubule-destabilizing kinesin XKCM1 regulates microtubule dynamic instability in cells.

Molecular biology of the cell ·Vol. 13 ·No. 8 ·2002-08-00 ·Pages 2718-31

Kline-Smith SL, Walczak CE

Abstract

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.

MeSH Terms
Animals Antibodies/metabolism Cell Line Immunohistochemistry Kinesins/genetics,metabolism Microinjections Microscopy, Fluorescence Microtubules/metabolism Mitosis/physiology Recombinant Fusion Proteins/genetics,metabolism Spindle Apparatus/metabolism Tubulin/metabolism Xenopus Proteins/genetics,metabolism
Chemicals
Antibodies KIF11 protein, Xenopus Recombinant Fusion Proteins Tubulin Xenopus Proteins Kinesins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Kline-Smith Susan L
Department of Anatomy and Cell Biology, Indiana University Medical Sciences Program, Bloomington 47405, USA.
Walczak Claire E
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
Published
2002-08-00
Pages
2718-31
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC117937
Subset
IM
Grants
NIGMS NIH HHS · R01 GM059618 · United States
NIGMS NIH HHS · GM-59618 · United States
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