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Changes in microtubule overlap length regulate kinesin-14-driven microtubule sliding

Nature Chemical Biology volume 13, pages 1245–1252 (2017)Cite this article

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Abstract

Microtubule-crosslinking motor proteins, which slide antiparallel microtubules, are required for the remodeling of microtubule networks. Hitherto, all microtubule-crosslinking motors have been shown to slide microtubules at a constant velocity until no overlap remains between them, leading to the breakdown of the initial microtubule geometry. Here, we show in vitro that the sliding velocity of microtubules, driven by human kinesin-14 HSET, decreases when microtubules start to slide apart, resulting in the maintenance of finite-length microtubule overlaps. We quantitatively explain this feedback using the local interaction kinetics of HSET with overlapping microtubules that cause retention of HSET in shortening overlaps. Consequently, the increased HSET density in the overlaps leads to a density-dependent decrease in sliding velocity and the generation of an entropic force that antagonizes the force exerted by the motors. Our results demonstrate that a spatial arrangement of microtubules can regulate the collective action of molecular motors through the local alteration of their individual interaction kinetics.

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Figure 1: HSET-driven microtubule sliding slows down when microtubules start to slide apart.
Figure 2: HSET-driven microtubule sliding slows down with increasing motor density.
Figure 3: Full-length GFP-HSET diffuses with different diffusion constants on single microtubules and in microtubule overlaps.
Figure 4: HSET confined in partial microtubule overlaps generates entropic forces.
Figure 5: Simulation of diffusible motors confined in a microtubule overlap explains the regulatory feedback by HSET.
Figure 6: Kinesin-14-driven microtubule sliding is regulated by changes in microtubule overlap length.

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Acknowledgements

We thank the members of the Diez laboratory for fruitful discussions, C. Walczak for the HSET plasmid DNA used as PCR template, G. Fink for helping to initiate the project and F. Ruhnow for generating the simulation kymographs. We acknowledge the financial support from the European Research Council (ERC starting grant 242933 to S.D.), the Deutsche Forschungsgemeinschaft (Heisenberg program grant DI 1226/4 and research unit SFG 877 grant DI 1226/5), the Czech Science Foundation (grant no. 15-17488S to Z.L. and 17-12496Y to M.B.), the Introduction of New Research Methods to BIOCEV (CZ.1.05/2.1.00/19.0390) project from the ERDF and institutional support from the Institute of Biotechnology RVO: 86652036. This work is part of the research program of the Foundation for Fundamental Research on Matter (FOM), which is part of the Netherlands Organization for Scientific Research (NWO).

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Author notes

  1. Marcus Braun and Zdenek Lansky: These authors contributed equally to this work.

Authors and Affiliations

  1. B CUBE Center for Molecular Bioengineering, Technische Universität Dresden, Dresden, Germany

    Marcus Braun, Zdenek Lansky, Agata Szuba, Friedrich W Schwarz, Aniruddha Mitra, Mengfei Gao, Annemarie Lüdecke & Stefan Diez

  2. Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany

    Marcus Braun, Zdenek Lansky, Agata Szuba, Friedrich W Schwarz, Aniruddha Mitra, Mengfei Gao, Annemarie Lüdecke & Stefan Diez

  3. Institute of Biotechnology CAS, BIOCEV, Vestec, Czech Republic

    Marcus Braun & Zdenek Lansky

  4. AMOLF, Amsterdam, The Netherlands

    Pieter Rein ten Wolde

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Contributions

M.B. and Z.L. conceived, performed and analyzed the experiments, generated the proteins, developed the mathematical model and wrote the manuscript. A.S., M.G. and A.L. performed and analyzed the sliding experiments, A.M. performed and analyzed the gliding experiments, F.W.S. performed the single-molecule diffusion analysis, P.R.T.W. developed the mathematical model and wrote the manuscript and S.D. conceived the experiments, developed the mathematical model and wrote the manuscript. All authors discussed the results and commented on the manuscript.

Corresponding authors

Correspondence to Pieter Rein ten Wolde or Stefan Diez.

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Braun, M., Lansky, Z., Szuba, A. et al. Changes in microtubule overlap length regulate kinesin-14-driven microtubule sliding. Nat Chem Biol 13, 1245–1252 (2017). https://doi.org/10.1038/nchembio.2495

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