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A mutant of the motor protein kinesin that moves in both directions on microtubules

Nature volume 406, pages 913–916 (2000)Cite this article

Abstract

Molecular motors move directionally to either the plus or the minus end of microtubules or actin filaments. Kinesin moves towards microtubule plus ends, whereas the kinesin-related Ncd motor moves to the minus ends. The ‘neck’—the region between the stalk and motor domain—is required for Ncd to move to microtubule minus ends1,2, but the mechanism underlying directional motor movement is not understood. Here we show that a single amino-acid change in the Ncd neck causes the motor to reverse directions and move with wild-type velocities towards the plus or minus end; thus, the neck is functional but directionality is defective. Mutation of a motor-core residue that touches the neck residue in crystal structures2,3 also results in movement in both directions, indicating that directed movement to the minus end requires interactions of the neck and motor core. Low-density laser-trap assays show that a conformational change or working stroke of the Ncd motor is directional and biased towards the minus end, whereas that of the neck mutant occurs in either direction. We conclude that the directional bias of the working stroke is dependent on neck/motor core interactions. Absence of these interactions removes directional constraints and permits movement in either direction.

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Figure 1: Neck and motor-core mutants.
Figure 2: Reversal and breakage of axoneme–microtubules in NK11 gliding assays.
Figure 3: Velocity of gliding on the NK11 mutant motor.
Figure 4: Low-density laser-trap assays of wild-type and mutant motors.

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Acknowledgements

We thank L. Yamaguchi and Y. Toyoshima for fluorescently labelled axoneme–microtubules, and J. Howard and W. Schief for valuable discussions. This work was supported by the NIH (S.A.E.), Japan Ministry of Education, Science, Sport & Culture (H.H.) and Human Frontier Science Program (S.A.E. and H.H.).

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Authors and Affiliations

  1. Department of Microbiology, Duke University Medical Center, Durham, 27710, North Carolina, USA

    Sharyn A. Endow

  2. Department of Metallurgy and Center for Interdisciplinary Research, Tohoku University, Sendai, 980-8579, Japan

    Hideo Higuchi

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Correspondence to Sharyn A. Endow.

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Endow, S., Higuchi, H. A mutant of the motor protein kinesin that moves in both directions on microtubules. Nature 406, 913–916 (2000). https://doi.org/10.1038/35022617

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