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Structure of a kinesin–tubulin complex and implications for kinesin motility

Abstract

The typical function of kinesins is to transport cargo along microtubules. Binding of ATP to microtubule-attached motile kinesins leads to cargo displacement. To better understand the nature of the conformational changes that lead to the power stroke that moves a kinesin's load along a microtubule, we determined the X-ray structure of human kinesin-1 bound to αβ-tubulin. The structure defines the mechanism of microtubule-stimulated ATP hydrolysis, which releases the kinesin motor domain from microtubules. It also reveals the structural linkages that connect the ATP nucleotide to the kinesin neck linker, a 15–amino acid segment C terminal to the catalytic core of the motor domain, to result in the power stroke. ATP binding to the microtubule-bound kinesin favors neck-linker docking. This biases the attachment of kinesin's second head in the direction of the movement, thus initiating each of the steps taken.

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Figure 1: Structure of the tubulin–kinesin complex.
Figure 2: The adjustment of the proteins of the tubulin–kinesin complex in the cryo-EM envelope of kinesin-1–decorated microtubules.
Figure 3: Conformational changes in tubulin-bound kinesin–ATP.
Figure 4: The kinesin-1 and Eg5 ATP-binding sites superpose well.
Figure 5: Neck-linker docking in tubulin-bound kinesin–ADP–AlF4.

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Acknowledgements

We are grateful to J. Skehel and L. Amos for comments on the manuscript. We thank I. Mignot for assistance and D. Mauchand (Unité Commune d'Expérimentation Animale, Institut National de la Recherche Agronomique, Jouy en Josas, France) for providing us with the material from which tubulin was purified. We also thank R. Vale (University of California, San Francisco, San Francisco, California, USA) for giving us the kinesin plasmid. Diffraction data were collected at the European Synchrotron Research Facility (ID29) and at SOLEIL synchrotron (PX1). We thank the machine and beamline groups for making these experiments possible. This work has also benefited from the facilities and expertise of IMAGIF (Centre de Recherche de Gif). We gratefully acknowledge support by the French Fondation pour la Recherche Médicale (DEQ20081213979 to M.K.), Fondation ARC pour la Recherche sur le Cancer (to B.G.) and Agence Nationale de la Recherche (grants ANR-09-BLAN-0071 and ANR-12-BSV8-0002-01 to B.G.) and the Chinese Ministry of Education (grant NCET-10-0604 to C.W.) and Science and Technology Commission of Shanghai Municipality (grant 11JC1413100 to C.W.).

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B.G., C.W. and M.K. designed research; B.G. crystallized the tubulin–DARPin–kinesin complex and determined its structure; W.W. and Q.J. characterized the kinesin-tubulin interaction biochemically; L.P., B.D. and A.P. provided the DARPin; B.G., C.W. and M.K. analyzed the data; B.G., C.W. and M.K. wrote the manuscript with input from all other authors.

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Correspondence to Chunguang Wang or Marcel Knossow.

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The authors declare no competing financial interests.

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Gigant, B., Wang, W., Dreier, B. et al. Structure of a kinesin–tubulin complex and implications for kinesin motility. Nat Struct Mol Biol 20, 1001–1007 (2013). https://doi.org/10.1038/nsmb.2624

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