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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

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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
Subjects: Q Science > QH Natural history
Q Science > QP Physiology
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
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:
DateEvent
12 July 2018Published
8 May 2018Accepted
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:
Project/Grant IDRIOXX Funder NameFunder ID
103895/Z/14/ZWellcome Trusthttp://dx.doi.org/10.13039/100010269
106151/Z/14/ZWellcome Trusthttp://dx.doi.org/10.13039/100010269
WM150020[RS] Royal Societyhttp://dx.doi.org/10.13039/501100000288
1090393[EPSRC] Engineering and Physical Sciences Research Councilhttp://dx.doi.org/10.13039/501100000266

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