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Dynein structure and power stroke

Nature volume 421, pages 715–718 (2003)Cite this article

Abstract

Dynein ATPases are microtubule motors that are critical to diverse processes such as vesicle transport and the beating of sperm tails; however, their mechanism of force generation is unknown. Each dynein comprises a head, from which a stalk and a stem emerge. Here we use electron microscopy and image processing to reveal new structural details of dynein c, an isoform from Chlamydomonas reinhardtii flagella, at the start and end of its power stroke. Both stem and stalk are flexible, and the stem connects to the head by means of a linker approximately 10 nm long that we propose lies across the head. With both ADP and vanadate bound, the stem and stalk emerge from the head 10 nm apart. However, without nucleotide they emerge much closer together owing to a change in linker orientation, and the coiled-coil stalk becomes stiffer. The net result is a shortening of the molecule coupled to an approximately 15-nm displacement of the tip of the stalk. These changes indicate a mechanism for the dynein power stroke.

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Figure 1: Negatively stained molecules of ADP•Vi-dynein c (left panel) and apo-dynein c (middle panel) from C. reinhardtii flagella.
Figure 2: Characteristic views of dynein c revealed by single-particle analysis.
Figure 3: Substructure and flexibility within left views of dynein c.
Figure 4: Structure and power stroke of dynein c.

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Acknowledgements

We thank J. Trinick for comments on an earlier draft. This work was supported in part by a NIH grant to J. Trinick and H. White, and by the BBSRC.

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Authors and Affiliations

  1. Astbury Centre for Structural Molecular Biology & School of Biomedical Sciences, University of Leeds, Leeds, LS2 9JT, UK

    Stan A. Burgess, Matt L. Walker & Peter J. Knight

  2. Kansai Advanced Research Centre, Communications Research Laboratory, Kobe, 651-2492, Japan

    Hitoshi Sakakibara & Kazuhiro Oiwa

Authors
  1. Stan A. Burgess
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Corresponding author

Correspondence to Stan A. Burgess.

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Supplementary information

These animations require the Shockwave plug in which can be downloaded at: http://www.macromedia.com/downloads/

Supplementary Movie 1 (ZIP 499 KB)

Conformations of the ADP.Vi-stalk. This movie shows a sequence of left-view image averages after classification of the stalk. When flexed (pointing left) the stalk is gently curved (clockwise from tip to base). When extended (pointing up) the stalk is straight.

Supplementary Movie 2 (ZIP 510 KB)

Comformations of the apo-stalk. This movie shows a sequence of left-view image averages after classification of the stalk. The stalk is straight except for a slight kink about 5 nm from the tip. It does not adopt the curved form seen in the flexed ADP.Vi-stalk, indicating a change in conformation of the stalk itself. As a result, the extent of flexibility (standard deviation of the distribution of stalk chord angles) is smaller compared to that in the ADP.Vi-stalk.

Supplementary Movie 3 (ZIP 493 KB)

Conformations of the ADP.Vi-stem. This movie shows a sequence of left-view image averages after classification of the stem. The narrow neck of the stem, close to the head, is the most flexible part of the ADP.Vi stem. There is little sign of extensibility of this region.

Supplementary Movie 4 (ZIP 492 KB)

Conformations of the apo-stem. This movie shows a sequence of left-view image averages after classification of the stem. The narrow neck of the stem, close to the head, is the most flexible part of the apo-stem. In general, the apo-neck lies closer to the head than the ADP.Vi-neck.

Supplementary Movie 5 (ZIP 1,896 KB)

Conformations of the stem and linker in undocked molecules. This movie shows a sequence of selected individual molecules, after alignment of their heads and is therefore more 'noisy'. The head adopts the same orientation as right views of apo-molecules (see Movie 6), but the undocked linker and stem emerge from the left side of the head.

Supplementary Movie 6 (ZIP 434 KB)

Conformations of the apo-stem in typical right views, i.e. with the linker docked onto the head. This movie shows a sequence of right-view image averages after classification of the stem. The stem emerges from the right side of the head, close to the stalk. The neck is not seen as clearly as in the left view, suggesting it is raised off the carbon substrate, compatible with the linker's ability to undock in this orientation.

Supplementary Movie 7 (ZIP 16 KB)

Power stroke of dynein. Left views of ADP.Vi- and apo-molecules in which the stem and stalk adopt the mean position relative to the head. Loss of ADP and Vi causes the apo-head to move towards the stem, and appear more ring-like. This displaces the tip of the stalk by 15 nm.

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Burgess, S., Walker, M., Sakakibara, H. et al. Dynein structure and power stroke. Nature 421, 715–718 (2003). https://doi.org/10.1038/nature01377

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