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Specific removal of TACC3-ch-TOG-clathrin at metaphase deregulates kinetochore fiber tension

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Cheeseman, L. P., Harry, Edward, McAinsh, Andrew D., Prior, I. A. and Royle, Stephen J. (2013) Specific removal of TACC3-ch-TOG-clathrin at metaphase deregulates kinetochore fiber tension. Journal of Cell Science, Volume 126 (Number 9). pp. 2102-2113. doi:10.1242/jcs.124834

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Official URL: http://dx.doi.org/10.1242/jcs.124834

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Abstract

Microtubule-associated proteins of the mitotic spindle are thought to be important for the initial assembly and the maintenance of spindle structure and function. However, distinguishing assembly and maintenance roles for a given protein is difficult. Most experimental methods for protein inactivation are slow and therefore affect both assembly and maintenance. Here, we have used ‘knocksideways’ to rapidly (∼5 minutes) and specifically remove TACC3–ch-TOG–clathrin non-motor complexes from kinetochore fibers (K-fibers). This method allows the complex to be inactivated at defined stages of mitosis. Removal of TACC3–ch-TOG–clathrin after nuclear envelope breakdown caused severe delays in chromosome alignment. Inactivation at metaphase, following a normal prometaphase, significantly delayed progression to anaphase. In these cells, K-fiber tension was reduced and the spindle checkpoint was not satisfied. Surprisingly, there was no significant loss of K-fiber microtubules, even after prolonged removal. TACC3–ch-TOG–clathrin removal during metaphase also resulted in a decrease in spindle length and significant alteration in kinetochore dynamics. Our results indicate that TACC3–ch-TOG–clathrin complexes are important for the maintenance of spindle structure and function as well as for initial spindle assembly.

Item Type: Journal Article
Divisions: Faculty of Science > Chemistry
Faculty of Science > Molecular Organisation and Assembly in Cells (MOAC)
Faculty of Medicine > Warwick Medical School
Journal or Publication Title: Journal of Cell Science
Publisher: The Company of Biologists Ltd.
ISSN: 0021-9533
Official Date: 2013
Dates:
DateEvent
2013Published
Volume: Volume 126
Number: Number 9
Page Range: pp. 2102-2113
DOI: 10.1242/jcs.124834
Status: Peer Reviewed
Publication Status: Published

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