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Inner-arm dynein c of Chlamydomonas flagella is a single-headed processive motor

Nature volume 400, pages 586–590 (1999)Cite this article

Abstract

Axonemal dyneins are force-generating ATPases that produce movement of eukaryotic cilia and flagella1,2. Several studies indicate that inner-arm dyneins mainly produce bending moments in flagella3,4 and that these motors have inherent oscillations in force and motility5,6,7,8. Processive motors such as kinesins have high duty ratios of attached to total ATPase cycle (attached plus detached) times9 compared to sliding motors such as myosin10. Here we provide evidence that subspecies-c, a single-headed axonemainner-arm dynein, is processive but has a low duty ratio. Ultrastructurally it is similar to other dyneins11,12,13,14, with a single globular head, long stem and a slender stalk that attaches to microtubules. In vitro studies of microtubules sliding over surfaces coated with subspecies-c at low densities (measured by single-molecule fluorescence) show that a single molecule is sufficient to move a microtubule more than 1 µm at 0.7 µm s−1. When many motors interact the velocity is 5.1 µm s−1, fitting a duty ratio of 0.14. Using optical trap nanometry, we show that beads carrying a single subspecies-c motor move processively along the microtubules in 8-nm steps but slip backwards under high loads. These results indicate that dynein subspecies-c functions in a very different way from conventional motor proteins, and has properties that could produce self-oscillation in vivo.

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Figure 1: Biochemical and ultrastructural characterization of subspecies-c.
Figure 2: Microtubule motion in vitro driven by subspecies-c.
Figure 3: Relationship between velocity of microtubule translation and subspecies-c surface density.
Figure 4: Optical trap nanometry of subspecies-c-coated beads.

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Acknowledgements

We thank M. Anson for discussion and comments on the manuscript and M.Kikumoto for single molecule measurements. This work was partly supported by the Hyogo Science and Technology Foundation (K.O. and H.K.) and a grant-in-aid from the Ministry of Education, Science and Culture of Japan (K.O.).

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Authors and Affiliations

  1. Kansai Advanced Research Center, Communications Research Laboratory, Kobe, 651-2401, Japan

    Hitoshi Sakakibara, Hiroaki Kojima, Yukako Sakai & Kazuhiro Oiwa

  2. Department of Fine Morphology, Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo, 108-8639, Japan

    Eisaku Katayama

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  1. Hitoshi Sakakibara
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Correspondence to Kazuhiro Oiwa.

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Sakakibara, H., Kojima, H., Sakai, Y. et al. Inner-arm dynein c of Chlamydomonas flagella is a single-headed processive motor. Nature 400, 586–590 (1999). https://doi.org/10.1038/23066

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