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bioRxiv · 10.1101/2025.01.31.635950

Multi-kinesin clusters impart mechanical stress that reveals mechanisms of microtubule breakage in cells

Geng, Q.·Bonilla, A.·Sandwith, S.·Verhey, K.
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Abstract

Microtubules are cytoskeletal filaments that provide structural support for numerous cellular processes. Despite their high rigidity, microtubules can be dramatically bent in cells and it is unknown how much force a microtubule can withstand before breaking. We find that liquid-liquid phase separation of the kinesin-3 motor KIF1C results in multi-kinesin clusters that entangle neighboring microtubules and impose a high level of mechanical stress that results in microtubule breakage and disassembly. Combining computational simulations and experiments, we show that microtubule fragmentation is enhanced by having a highly processive kinesin motor domain, a stiff clustering mechanism, and sufficient drag force on the microtubules. We estimate a rupture force for microtubules in cells of 70-120 pN, which is lower than previous estimates based on in vitro studies with taxol-stabilized microtubules. These results indicate that the presence of multiple kinesins on a cargo has the potential to cause microtubule breakage. We propose that mechanisms exist to protect microtubule integrity by releasing either the motor-cargo or motor-microtubule interaction, thereby preventing the accumulation of mechanical stress upon the engagement of multi-motor clusters with microtubules.

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Geng, Q., Bonilla, A., Sandwith, S., Verhey, K.. 2025-02-03. Multi-kinesin clusters impart mechanical stress that reveals mechanisms of microtubule breakage in cells. https://doi.org/10.1101/2025.01.31.635950

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