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An extended microtubule-binding structure within the dynein motor domain

Nature volume 390, pages 636–639 (1997)Cite this article

Abstract

Flagellar dynein was discovered over 30 years ago as the first motor protein capable of generating force along microtubules1. A cytoplasmic form of dynein has also been identified which is involved in mitosis and a wide range of other intracellular movements2 (reviewed in ref. 3). Rapid progress has been made on understanding the mechanism of force production by kinesins and myosins4,5,6,7,8. In contrast, progress in understanding the dyneins has been limited by their great size (relative molecular mass 1,000K–2,000K) and subunit complexity. We now report evidence that the entire carboxy-terminal two-thirds of the 532K force-producing heavy chain subunit is required for ATP-binding activity. We further identify a microtubule-binding domain, which, surprisingly, lies well downstream of the entire ATPase region and is predicted to form a hairpin-like stalk. Direct ultrastructural analysis of a recombinant fragment confirms this model, and suggests that the mechanism for dynein force production differs substantially from that of other motor proteins.

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Figure 1: Effect of a dynein heavy-chain point mutation on VO4-mediated photocleavage.
Figure 2: Summary of recombinant dynein heavy-chain behaviour in VO4 photocleavage and microtubule binding assays.
Figure 3: Co-distribution of mutant dynein heavy chain with microtubules.
Figure 5: Structural and functional predictions for the dyein stalk.
Figure 4: Microtubule-binding and electron microscopic analysis of recombinant microtubule binding domain.

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Acknowledgements

We thank A. Mikami, P. Okamoto, S. Hughes, C. Echeverri, E. Luna, C. Wilkerson and U. Goodenough for discussions; P. McNulty for assistance in the preparation of this manuscript; R. Roth for technical assistance; M. Morgan for computer graphics for the EM images; E. Steuer for cytoplasmic dynein; and U. Goodenough for axonemal dynein.

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Authors and Affiliations

  1. Worcester Foundation for Biomedical Research, 222 Maple Avenue, Shrewsbury, 01545, Massachusetts

    Melissa A. Gee & Richard B. Vallee

  2. University of Massachusetts Medical Center, Worcester, 01655, Massachusetts, USA

    Melissa A. Gee & Richard B. Vallee

  3. Washington University, 660 South Euclid Avenue, St. Louis, 63110, Missouri, USA

    John E. Heuser

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  1. Melissa A. Gee
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Correspondence to Richard B. Vallee.

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Gee, M., Heuser, J. & Vallee, R. An extended microtubule-binding structure within the dynein motor domain. Nature 390, 636–639 (1997). https://doi.org/10.1038/37663

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