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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
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 | ||||
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| Divisions: | Faculty of Science > Chemistry Faculty of Science > Molecular Organisation and Assembly in Cells (MOAC) Faculty of Medicine > Warwick Medical School |
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| Journal or Publication Title: | Journal of Cell Science | ||||
| Publisher: | The Company of Biologists Ltd. | ||||
| ISSN: | 0021-9533 | ||||
| Official Date: | 2013 | ||||
| Dates: |
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| 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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