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Phospholipid-dependent regulation of the motor activity of myosin X

Nature Structural & Molecular Biology volume 18, pages 783–788 (2011)Cite this article

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Abstract

Myosin X is involved in the reorganization of the actin cytoskeleton and protrusion of filopodia. Here we studied the molecular mechanism by which bovine myosin X is regulated. The globular tail domain inhibited the motor activity of myosin X in a Ca2+-independent manner. Structural analysis revealed that myosin X is monomeric and that the band 4.1-ezrin-radixin-moesin (FERM) and pleckstrin homology (PH) domains bind to the head intramolecularly, forming an inhibited conformation. Binding of phosphatidylinositol-3,4,5-triphosphate (PtdIns(3,4,5)P3) to the PH domain reversed the tail-induced inhibition and induced the formation of myosin X dimers. Consistently, disruption of the binding of PtdIns(3,4,5)P3 attenuated the translocation of myosin X to filopodial tips in cells. We propose the following mechanism: first, the tail inhibits the motor activity of myosin X by intramolecular head-tail interactions to form the folded conformation; second, phospholipid binding reverses the inhibition and disrupts the folded conformation, which induces dimer formation, thereby activating the mechanical and cargo transporter activity of myosin X.

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Figure 1: Schematic diagram of myosin X constructs used in this study.
Figure 2: The actin-activated ATPase activity of M10full and M10ΔGTD.
Figure 3: Electron microscopy analysis of myosin X constructs.
Figure 4: Binding of PtdIns(3,4,5)P3 to myosin X abolishes the tail-induced inhibition of the actin-activated ATPase activity of myosin X.
Figure 5: Activation of the actin-gliding velocity of M10full by PtdIns(3,4,5)P3.
Figure 6: Induction of myosin X dimer formation by PtdIns(3,4,5)P3 measured by cross-linking.
Figure 7: Phospholipid binding is crucial for the filopodial tip localization of myosin X.

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Acknowledgements

We thank R. Fenton (University of Massachusetts) for reading the manuscript and K. Homma for the parent construct of myosin X. This work was supported by US National Institutes of Health (NIH) grants DC006103, AR048526, AR048898 and HL073050 to M.I. and Korea Basic Science Institute grant T31760 to H.S.J.

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  1. Nobuhisa Umeki, Hyun Suk Jung and Tsuyoshi Sakai: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, Massachusetts, USA

    Nobuhisa Umeki, Tsuyoshi Sakai, Osamu Sato, Reiko Ikebe & Mitsuo Ikebe

  2. Division of Electron Microscopic Research, Korea Basic Science Institute, Daejeon, Korea

    Hyun Suk Jung

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  1. Nobuhisa Umeki
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Contributions

N.U. performed the ATPase assays of myosin X constructs, cross-linking assay and prepared myosin X proteins for EM experiments. H.S.J. performed EM image analysis and wrote the EM part of the paper. T.S. performed lipid binding assay, cross-linking, cell imaging, measurement of the activity of PH domain mutants and production of some myosin X constructs. O.S. performed the actin gliding assay of myosin X. R.I. prepared myosin X expression constructs. M.I. supervised the whole project and wrote the paper.

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Correspondence to Mitsuo Ikebe.

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The authors declare no competing financial interests.

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Supplementary Figures 1–8 and Supplementary Methods (PDF 5877 kb)

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Umeki, N., Jung, H., Sakai, T. et al. Phospholipid-dependent regulation of the motor activity of myosin X. Nat Struct Mol Biol 18, 783–788 (2011). https://doi.org/10.1038/nsmb.2065

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