Letter | Published:

Autoinhibition and cooperative activation mechanisms of cytoplasmic dynein

Nature Cell Biology volume 16, pages 11181124 (2014) | Download Citation

Abstract

Cytoplasmic dynein is a two-headed microtubule-based motor responsible for diverse intracellular movements, including minus-end-directed transport of organelles1,2,3. The motility of cargo transporters is regulated according to the presence or absence of cargo4,5,6; however, it remains unclear how cytoplasmic dynein achieves such regulation. Here, using a recombinant and native dynein complex in vitro, we show that lone, single dynein molecules are in an autoinhibited state, in which the two motor heads are stacked together. In this state, dynein moves diffusively along a microtubule with only a small bias towards the minus end of the microtubule. When the two heads were physically separated by a rigid rod, the movement of dynein molecules became directed and processive. Furthermore, assembly of multiple dynein molecules on a single cargo enabled them to move unidirectionally and generate force cooperatively. We thus propose a mechanism of autonomous on–off switching of cargo transport, in which single dynein molecules in the cell are autoinhibited through intramolecular head–head stacking and become active when they assemble as a team on a cargo.

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Acknowledgements

We thank K. H. Bui, T. Ishikawa and H. Sakakibara for technical instruction in electron microscopy, K. Sutoh for critical comments on the manuscript, K. Kawaguchi for helpful discussion, K. Yamamoto for early work on DNA nanotubes, M. Amino and Y. Watari for technical assistance, K. Shibata for constructing the monomeric yeast dynein construct and Y. Utsumi for the chicken α-actinin plasmid. This work was supported by a Grant-in-Aid for the Japan Society for the Promotion of Science (JSPS) fellowship from JSPS, a JSPS Grant-in-Aid for Scientific Research (B), a JSPS Grant-in-Aid for Scientific Research (C) and a Grant-in-Aid for Young Scientists (B) from the Ministry of Education, Culture, Sports, Science and Technology, Japan.

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Affiliations

  1. Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Meguro-ku, Tokyo 153-8902, Japan

    • Takayuki Torisawa
    • , Muneyoshi Ichikawa
    • , Kei Saito
    •  & Yoko Y. Toyoshima
  2. Advanced ICT Research Institute, National Institute of Information and Communications Technology, Kobe, Hyogo 651-2492, Japan

    • Takayuki Torisawa
    • , Akane Furuta
    • , Kazuhiro Oiwa
    • , Hiroaki Kojima
    •  & Ken’ya Furuta
  3. CREST, Japan Science and Technology Agency, Chiyoda-ku, Tokyo 102-0076, Japan

    • Takayuki Torisawa
    •  & Kazuhiro Oiwa
  4. Graduate School of Life Science, University of Hyogo, Harima Science Park City, Hyogo 678-1297, Japan

    • Kazuhiro Oiwa

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Contributions

T.T., Y.Y.T. and K.F. designed research, T.T., M.I., A.F., K.S. and K.F. performed experiments and T.T., M.I., K.O., H.K., Y.Y.T. and K.F. analysed data and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Yoko Y. Toyoshima or Ken’ya Furuta.

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Videos

  1. 1.

    Diffusive movement of single full-length dynein molecules (3 × real-time).

    The movements were observed in the presence of 1 mM ATP. Green lines indicate the position of the MT. Bars represent 3 μm.

  2. 2.

    Directed movement of single ACTN-D382 dynein molecules (10 × real-time).

    The movements were observed in the presence of 1 mM ATP. Bars represent 3 μm.

  3. 3.

    Directed movement of single GST-D425-GFP dynein molecules (10 × real-time).

    The movements were observed in the presence of 1 mM ATP. Bars represent 3 μm.

  4. 4.

    Directed movement of two coupled full-length dynein molecules (10 × real-time).

    The movements were observed in the presence of 1 mM ATP. Bars represent 3 μm.

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DOI

https://doi.org/10.1038/ncb3048

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