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Kif15 functions as an active mechanical ratchet
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McHugh, Toni, Drechsler, Hauke, McAinsh, Andrew D., Carter, Nicholas J. and Cross, R. A. (2018) Kif15 functions as an active mechanical ratchet. Molecular Biology of the Cell, 29 (4). pp. 1743-1752. doi:10.1091/mbc.E18-03-0151 ISSN 1059-1524.
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Official URL: https://doi.org/10.1091/mbc.E18-03-0151
Abstract
Kif15 is a kinesin-12 that contributes critically to bipolar spindle assembly in humans. Here we use force-ramp experiments in an optical trap to probe the mechanics of single Kif15 molecules under hindering or assisting loads and in a variety of nucleotide states. Whilst unloaded Kif15 is established to be highly processive, we find that under hindering loads, Kif15 takes <∼10 steps. As hindering load is increased, Kif15 forestep:backstep ratio decreases exponentially, with stall occurring at 6 pN. By contrast, under assisting loads, Kif15 detaches readily and rapidly, even from its AMPPNP state. Kif15 mechanics thus depend markedly on the loading direction. Kif15 interacts with a binding partner, Tpx2, and we show that Tpx2 locks Kif15 to microtubules under both hindering and assisting loads. Overall, our data predict that Kif15 in the central spindle will act as a mechanical ratchet, supporting spindle extension but resisting spindle compression.
| Item Type: | Journal Article | |||||||||||||||
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| Subjects: | Q Science > QH Natural history Q Science > QP Physiology |
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| Divisions: | Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School > Biomedical Sciences > Cell & Developmental Biology Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School > Biomedical Sciences Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School |
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| Library of Congress Subject Headings (LCSH): | Kinesin, Spindle (Cell division), Nucleotides, Microtubules | |||||||||||||||
| Journal or Publication Title: | Molecular Biology of the Cell | |||||||||||||||
| Publisher: | American Society for Cell Biology | |||||||||||||||
| ISSN: | 1059-1524 | |||||||||||||||
| Official Date: | 12 July 2018 | |||||||||||||||
| Dates: |
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| Volume: | 29 | |||||||||||||||
| Number: | 4 | |||||||||||||||
| Page Range: | pp. 1743-1752 | |||||||||||||||
| DOI: | 10.1091/mbc.E18-03-0151 | |||||||||||||||
| Status: | Peer Reviewed | |||||||||||||||
| Publication Status: | Published | |||||||||||||||
| Access rights to Published version: | Restricted or Subscription Access | |||||||||||||||
| Date of first compliant deposit: | 24 May 2018 | |||||||||||||||
| Date of first compliant Open Access: | 18 October 2018 | |||||||||||||||
| RIOXX Funder/Project Grant: |
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