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A lever-arm rotation drives motility of the minus-end-directed kinesin Ncd

Nature volume 439, pages 875–878 (2006)Cite this article

Abstract

Kinesins are microtubule-based motor proteins that power intracellular transport1,2. Most kinesin motors, exemplified by Kinesin-1, move towards the microtubule plus end, and the structural changes that govern this directional preference have been described3,4,5. By contrast, the nature and timing of the structural changes underlying the minus-end-directed motility of Kinesin-14 motors (such as Drosophila Ncd6,7) are less well understood. Using cryo-electron microscopy, here we demonstrate that a coiled-coil mechanical element of microtubule-bound Ncd rotates ∼70° towards the minus end upon ATP binding. Extending or shortening this coiled coil increases or decreases velocity, respectively, without affecting ATPase activity. An unusual Ncd mutant that lacks directional preference8 shows unstable nucleotide-dependent conformations of its coiled coil, underscoring the role of this mechanical element in motility. These results show that the force-producing conformational change in Ncd occurs on ATP binding, as in other kinesins, but involves the swing of a lever-arm mechanical element similar to that described for myosins.

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Figure 1: 3D maps of Ncd–microtubule complexes by cryo-EM.
Figure 2: Ncd mutants with truncated or extended necks.
Figure 3: Model of the Ncd motility cycle.

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Acknowledgements

We thank S. Rice, J. Kardon and J. Ebstein for initial work on this project. This work was supported by NIH grants (to R.D.V. and R.A.M.). Some of the work presented here was conducted at the National Resource for Automated Molecular Microscopy, which is supported by the NIH.

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Authors and Affiliations

  1. The Howard Hughes Medical Institute, and the Department of Cellular and Molecular Pharmacology, University of California San Francisco, 600 16th Street, California, 94107, San Francisco, USA

    Nicholas F. Endres & Ronald D. Vale

  2. Center for Integrative Molecular Biosciences, Department of Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, California, 92037, La Jolla, USA

    Craig Yoshioka & Ronald A. Milligan

Authors
  1. Nicholas F. Endres
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  2. Craig Yoshioka
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  3. Ronald A. Milligan
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  4. Ronald D. Vale
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Correspondence to Ronald D. Vale.

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Supplementary information

Supplementary Figure Legends

Text to accompany Supplementary Figures 1–5 referred to in the text, with additional references. (DOC 39 kb)

Supplementary Figure 1

This figure demonstrates that the Ncd neck position is not determined by the apparent interaction between the tip of the neck and the adjacent motor domain. (JPG 174 kb)

Supplementary Figure 2

This figure demonstrates that the Ncd neck occupies a similar position in the AMPPNP and ADPAlF4- states. (JPG 175 kb)

Supplementary Figure 3

This figure shows microtubule binding interface implied by docking experiments are reasonable. (JPG 140 kb)

Supplementary Figure 4

Statistical different maps provide additional support for our model for the bidirectional motility of N340K, and demonstrate that the AMPPNP and ADP-ALF4- states for this mutant are the same. (JPG 196 kb)

Supplementary Figure 5

Gel Filtration results which confirm the appropriate size for the Ncd heterodimer and monomer. (PDF 220 kb)

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Endres, N., Yoshioka, C., Milligan, R. et al. A lever-arm rotation drives motility of the minus-end-directed kinesin Ncd. Nature 439, 875–878 (2006). https://doi.org/10.1038/nature04320

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