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ATP hydrolysis cycle–dependent tail motions in cytoplasmic dynein

Nature Structural & Molecular Biology volume 12, pages 513–519 (2005)Cite this article

Abstract

The motor protein dynein is predicted to move the tail domain, a slender rod-like structure, relative to the catalytic head domain to carry out its power stroke. Here, we investigated ATP hydrolysis cycle–dependent conformational dynamics of dynein using fluorescence resonance energy transfer analysis of the dynein motor domain labeled with two fluorescent proteins. We show that dynein adopts at least two conformational states (states I and II), and the tail undergoes ATP-induced motions relative to the head domain during transitions between the two states. Our measurements also suggest that in the course of the ATP hydrolysis cycle of dynein, the tail motion from state I to state II takes place in the ATP-bound state, whereas the motion from state II to state I occurs in the ADP-bound state. The latter tail motion may correspond to the predicted power stroke of dynein.

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Figure 1: Design of FRET-based sensors for detection of conformational states of dynein.
Figure 2: Detection of ATP-induced conformational changes in the dynein motor domain by means of FRET measurements.
Figure 3: Effect of nucleotides on the FRET efficiency between the BFP and GFP moieties within HG380B2 dynein.
Figure 4: Effect of ATPase active site mutations on nucleotide-induced changes in FRET within the HFG380B2 dynein.
Figure 5: Model of the ATP hydrolysis cycle–dependent tail motions in the dynein motor domain.

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Acknowledgements

We thank P.J. Van Haastert for providing us with the tetracycline-regulated expression system. This work was partly supported by the Grant-in-Aid for Scientific Research (B) from the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT), by the Grant-in-Aid for Scientific Research on Priority Areas from MEXT, by the Special Coordination Funds for Promoting Science and Technology from MEXT and by the Core Research for Evolutional Science and Technology (CREST) program grant from the Japan Science and Technology Agency (JST).

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  1. Takahide Kon and Toshifumi Mogami: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Life Sciences, Graduate School of Arts and Sciences, University of Tokyo, Komaba 3-8-1, Tokyo, 153-8902, Japan

    Takahide Kon, Toshifumi Mogami, Reiko Ohkura, Masaya Nishiura & Kazuo Sutoh

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  1. Takahide Kon
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Correspondence to Kazuo Sutoh.

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Kon, T., Mogami, T., Ohkura, R. et al. ATP hydrolysis cycle–dependent tail motions in cytoplasmic dynein. Nat Struct Mol Biol 12, 513–519 (2005). https://doi.org/10.1038/nsmb930

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