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J Biol Chem, Vol. 275, Issue 16, 12346-12352, April 21, 2000
From the Department of Biochemistry, Dartmouth Medical School,
Hanover, New Hampshire 03755
Microtubules induced to polymerize with taxol in
a mammalian mitotic extract organize into aster-like arrays in a
centrosome-independent process that is driven by microtubule motors and
structural proteins. These microtubule asters accurately reflect the
noncentrosomal aspects of mitotic spindle pole formation. We show here
that colonic-hepatic tumor-overexpressed gene (ch-TOGp)
is an abundant component of these asters. We have prepared
ch-TOGp-specific antibodies and show by immunodepletion that ch-TOGp is
required for microtubule aster assembly. Microtubule polymerization is
severely inhibited in the absence of ch-TOGp, and silver stain analysis
of the ch-TOGp immunoprecipitate indicates that it is not present in a
preformed complex and is the only protein removed from the extract
during immunodepletion. Furthermore, the reduction in microtubule
polymerization efficiency in the absence of ch-TOGp is dependent on
ATP. These results demonstrate that ch-TOGp is a major constituent of
microtubule asters assembled in a mammalian mitotic extract and that it
is required for robust microtubule polymerization in an
ATP-dependent manner in this system even though taxol is
present. These data, coupled with biochemical and genetic data derived
from analysis of ch-TOGp-related proteins in other organisms, indicate
that ch-TOGp is a key factor regulating microtubule dynamics during mitosis.
ch-TOGp Is Required for Microtubule Aster Formation in a
Mammalian Mitotic Extract*
*
This work was supported by Grant GM51542 from the National
Institutes of Health.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed. Tel.: 603-650-1990;
Fax: 603-650-1128; E-mail: duane.a.compton@dartmouth.edu.
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