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Research Article |
1 Department of Zoology, University of Wisconsin, Madison, Madison, WI 53706,
USA
2 Program in Cellular and Molecular Biology, University of Wisconsin, Madison,
Madison, WI 53706, USA
* Author for correspondence (e-mail: wmbement{at}facstaff.wisc.edu )
Accepted 9 January 2002
The microtubule, F-actin, and intermediate filament systems are often
studied as isolated systems, yet the three display mutual interdependence in
living cells. To overcome limitations inherent in analysis of polymer-polymer
interactions in intact cells, associations between these systems were assessed
in Xenopus egg extracts. In both fixed and unfixed extract
preparations, cytokeratin associated with F-actin cables that spontaneously
assembled in the extracts. Time-course experiments revealed that at early time
points cytokeratin cables were invariably associated with F-actin cables,
while at later time points they could be found without associated F-actin. In
extract samples where F-actin assembly was prevented, cytokeratin formed
unorganized aggregates rather than cables. Dynamic imaging revealed transport
of cytokeratin by moving F-actin as well as examples of cytokeratin release
from F-actin. Experimental alteration of F-actin network organization by
addition of 
-actinin resulted in a corresponding change in the
organization of the cytokeratin network. Finally, pharmacological disruption
of the F-actin network in intact, activated eggs disrupted the normal pattern
of cytokeratin assembly. These results provide direct evidence for an
association between F-actin and cytokeratin in vitro and in vivo, and indicate
that this interaction is necessary for proper cytokeratin assembly after
transition into the first mitotic interphase of Xenopus.
Key words: Cytoskeleton interactions, Actin, microtubules, Intermediate filaments
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