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A torque component in the kinesin-1 power stroke

Nature Chemical Biology volume 1, pages 338–341 (2005)Cite this article

Abstract

Kinesin-1 is a twin-headed molecular motor that moves along microtubules in 8-nm steps, using a walking action in which the two heads interact alternately with the microtubule1,2,3,4. Constructs with only one head can also produce impulses of force and motion5,6,7, indicating that the walking action is an amplification strategy that leverages an underlying force-generating event. Recent work suggests that directional force is produced either by directionally biased selection of microtubule binding sites8,9 or by a conformational change subsequent to the binding event10,11,12. We report here that surface-attached rat kinesin-1 monomers drive counterclockwise rotation of sliding microtubules around their axes, and that by manipulating the assay geometry, we could reduce or block the torsional motion with negligible effects on the axial motion. We can account for this behavior on the simple assumption that kinesin heads tend to bind to the closest available tubulin heterodimer in the lattice, but only in the case where an additional biasing process is present that shifts the start position for diffusion-to-capture toward the microtubule plus end by ∼1 nm.

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Figure 1: Single-headed kinesins induce microtubules to rotate while sliding.
Figure 2: Axial and rotational microtubule velocities.
Figure 3: Mechanochemical models for kinesin-1 heads.
Figure 4: The surface lattice of kinesin-binding sites on microtubules.

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Acknowledgements

We thank D. Drummond, N. Carter, P. Gross and T. Oelgeschlager for advice. Supported by Marie Curie Cancer Care, the Japan Society for the Promotion of Science and the Oxford IRC in Bionanotechnology.

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Authors and Affiliations

  1. Molecular Motors Group, Marie Curie Research Institute, The Chart, Oxted, RH8 0TL, Surrey, UK

    Junichiro Yajima & Robert A Cross

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Correspondence to Robert A Cross.

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Supplementary information

Supplementary Fig. 1

RK340-Sp1 binding to DNA (a) Gel mobility shift assay of RK340-Sp1 binding to an 80 bp double stranded DNA that included the Sp1 GC box recognition sequence (0, 1, 5, 10, 20, 40, 70, and 100 nM proteins in lane 0-7, respectively). (b) Apparent equilibrium binding curve obtained by calculating the fraction of DNA bound at varying RK340-Sp1 concentrations. The binding constant was determined by fitting to the equation Øb = [protein] / ([protein] + Kd) to obtain the dissociation constant (Kd). The observed dissociation constant was Kd = 21 ± 6 nM, which was close to the value as reported previously [Richard W. et al. (1992) PNAS 89 9759-63]. (PDF 588 kb)

Supplementary Video 1

Microtubule rotation driven by RK340Gel. The scale bar is 2 μm. The time code is hours : minutes : seconds : centiseconds. (MOV 4247 kb)

Supplementary Video 2

Sliding of a microtubule blocked in rotation. The arrow moving south-north marks a microtubule with a long side-arm that is unable to rotate. Sliding continues (see text) at close to wild type velocity. The arrow moving west-east tracks a microtubule with a short side-arm that is rotating normally. Time code and scale bar as in Supplementary Video 1. (MOV 2939 kb)

Supplementary Video 3

Microtubule rotation driven by RK345L-Sp1-biotinylated DNA. Time code and scale bar as in Supplementary Video 1. (MOV 4240 kb)

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Yajima, J., Cross, R. A torque component in the kinesin-1 power stroke. Nat Chem Biol 1, 338–341 (2005). https://doi.org/10.1038/nchembio740

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