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Myosin-V makes two brownian 90° rotations per 36-nm step

Nature Structural & Molecular Biology volume 14, pages 968–973 (2007)Cite this article

Abstract

Myosin-V processively walks on actin filaments in a hand-over-hand fashion. The identical structures of the heads predict a symmetric hand-over-hand mechanism where regular, unidirectional rotation occurs during a 36-nm step. We investigated this by observing how fixed myosin-V rotates actin filaments. Actin filaments randomly rotated 90° both clockwise and counter-clockwise during each step. Furthermore, ATP-dependent rotations were regularly followed by ATP-independent ones. Kinetic analysis indicated that the two 90° rotations relate to the coordinated unbinding and rebinding of the heads with actin. We propose a 'brownian rotation hand-over-hand' model, in which myosin-V randomly rotates by thermally twisting its elastic neck domains during the 36-nm step. The brownian rotation may be advantageous for cargo transport through a crowded actin meshwork and for carrying cargoes reliably via multiple myosin-V molecules in the cell.

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Figure 1: Observation of actin-filament rotation by single myosin-V molecules.
Figure 2: 90° rotation of actin filaments.
Figure 3: Dwell times of 90° rotation steps.
Figure 4: Translational movement of actin filaments by single myosin-V molecules.
Figure 5: Correlation of the 90° rotation directions.
Figure 6: A brownian rotational hand-over-hand model.

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Acknowledgements

This work was supported by Core Research for Evolutional Science and Technology program of the Japanese Science and Technology Corporation and the Special Coordination Funds for Promoting Science and Technology: Yuragi Project of the Ministry of Education, Culture, Sports, Science and Technology, Japan. We are grateful to O. Ohara and H. Yamakawa at the Kazusa DNA Research Institute for providing complementary DNA fragments encoding human myosin-V; T. Wazawa and H. Tanaka for technical support in the single-molecule assay; T. Mimuro-Ichinose for generating the human myosin-V construct; Y. Ishii and P. Karagiannis for critically reading the manuscript and for helpful discussions; and colleagues of the Yanagida laboratory at the Single Molecule Processes Project, Formation of Soft Nanomachines, and Osaka University for discussion.

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Authors and Affiliations

  1. Laboratories for Nanobiology, Graduate School of Frontier Biosciences, Osaka University, 1-3, Yamadaoka, Suita, Osaka, 565-0871, Japan

    Yasunori Komori, Atsuko H Iwane & Toshio Yanagida

  2. Formation of Soft Nano-Machines, Core Research for Evolutional Science and Technology, Japanese Science and Technology Corporation, 1-3, Yamadaoka, Suita, Osaka, 565-0871, Japan

    Yasunori Komori & Toshio Yanagida

  3. Department of Physiology and Biosignaling, Graduate School of Medicine, Osaka University, 2-2, Yamadaoka, Suita, Osaka, 565-0871, Japan

    Atsuko H Iwane & Toshio Yanagida

  4. Special Coordination Funds for Promoting Science and Technology: Yuragi Project of the Ministry of Education, Culture, Sports, Science and Technology, 2-5-1, Marunouchi, Chiyoda-ku, Tokyo, 100-8959, Japan

    Toshio Yanagida

Authors
  1. Yasunori Komori
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Contributions

Y.K. conceived the experiments, constructed the microscopy system, devised custom software, prepared materials, performed experiments and analyzed data. A.H.I. designed, constructed and expressed recombinant myosin-V. All processes were supervised by T.Y. Y.K., A.H.I. and T.Y. wrote the manuscript.

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Correspondence to Toshio Yanagida.

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Supplementary Figures 1–4, Supplementary Discussion (PDF 472 kb)

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Komori, Y., Iwane, A. & Yanagida, T. Myosin-V makes two brownian 90° rotations per 36-nm step. Nat Struct Mol Biol 14, 968–973 (2007). https://doi.org/10.1038/nsmb1298

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