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Probing microtubule polymerisation state at single kinetochores during metaphase chromosome motion
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Armond, Jonathan W., Vladimirou, Elina, Erent, Muriel, McAinsh, Andrew D. and Burroughs, Nigel John (2015) Probing microtubule polymerisation state at single kinetochores during metaphase chromosome motion. Journal of Cell Science, Volume 128 (Number 10). pp. 1991-2001. doi:10.1242/jcs.168682 ISSN 0021-9533.
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Official URL: http://dx.doi.org/10.1242/jcs.168682
Abstract
Kinetochores regulate the dynamics of attached microtubule bundles (kinetochore-fibres, K-fibres) to generate the forces necessary for chromosome movements in mitosis. Current models suggest that poleward-moving kinetochores are attached to depolymerising K-fibres and anti-poleward-moving kinetochores to polymerising K-fibres. How the dynamics of individual microtubules within the K-fibre relate to poleward and anti-poleward movements is poorly understood. To investigate this, we developed a live-cell imaging assay combined with computational image analysis that allows eGFP-tagged EB3 (also known as MAPRE3) to be quantified at thousands of individual metaphase kinetochores as they undergo poleward and anti-poleward motion. Surprisingly, we found that K-fibres are incoherent, containing both polymerising and depolymerising microtubules – with a small polymerisation bias for anti-poleward-moving kinetochores. K-fibres also display bursts of EB3 intensity, predominantly on anti-poleward-moving kinetochores, equivalent to more coherent polymerisation, and this was associated with more regular oscillations. The frequency of bursts and the polymerisation bias decreased upon loss of kinesin-13, whereas loss of kinesin-8 elevated polymerisation bias. Thus, kinetochores actively set the balance of microtubule polymerisation dynamics in the K-fibre while remaining largely robust to fluctuations in microtubule polymerisation.
| Item Type: | Journal Article | ||||||||||
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| Subjects: | Q Science > QH Natural history > QH301 Biology | ||||||||||
| Divisions: | Faculty of Science, Engineering and Medicine > Science > Mathematics Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School > Biomedical Sciences > Microbiology & Infection Faculty of Science, Engineering and Medicine > Research Centres > Warwick Systems Biology Centre Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School |
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| Library of Congress Subject Headings (LCSH): | Cytology—Research, Cell division, Mitosis | ||||||||||
| Journal or Publication Title: | Journal of Cell Science | ||||||||||
| Publisher: | The Company of Biologists Ltd. | ||||||||||
| ISSN: | 0021-9533 | ||||||||||
| Official Date: | 15 May 2015 | ||||||||||
| Dates: |
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| Volume: | Volume 128 | ||||||||||
| Number: | Number 10 | ||||||||||
| Page Range: | pp. 1991-2001 | ||||||||||
| DOI: | 10.1242/jcs.168682 | ||||||||||
| Status: | Peer Reviewed | ||||||||||
| Publication Status: | Published | ||||||||||
| Access rights to Published version: | Open Access (Creative Commons) | ||||||||||
| Date of first compliant deposit: | 16 March 2016 | ||||||||||
| Date of first compliant Open Access: | 16 March 2016 | ||||||||||
| Funder: | Biotechnology and Biological Sciences Research Council (Great Britain) (BBSRC) | ||||||||||
| Grant number: | BB/I021353/1 | ||||||||||
| Related URLs: |
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