Goulet, Adeline and Major, J. and Jung, Y. and Grossmann, S.P. and Rosenfeld, S.S. and Moores, Carolyn A. (2014) Comprehensive structural model of the mechanochemical cycle of a mitotic motor highlights molecular adaptations in the kinesin family. Proceedings of the National Academy of Sciences of the United States of America 111 (5), pp. 1837-1842. ISSN 0027-8424.
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Abstract
Kinesins are responsible for a wide variety of microtubule-based, ATP-dependent functions. Their motor domain drives these activities but the molecular adaptations that specify these diverse and essential cellular activities are poorly understood. It has been assumed that the first identified kinesin - the transport motor kinesin-1 – is the mechanistic paradigm for the entire superfamily, but accumulating evidence suggests that this is not the case. To address the deficits in our understanding of the molecular basis of functional divergence within the kinesin superfamily, we studied kinesin-5s, which are essential mitotic motors whose inhibition blocks cell division. Using cryo-electron microscopy and subnanometer resolution structure determination, we have visualised conformations of microtubule-bound human kinesin-5 motor domain at successive steps in its ATPase cycle. Following ATP hydrolysis, nucleotide-dependent conformational changes in the active site are allosterically propagated into rotations of the motor domain and uncurling of the drugbinding loop L5. In addition, the mechanical neck-linker element that is crucial for motor stepping undergoes discrete, ordered displacements. We also observed large reorientations of the motor N-terminus that indicate its importance for kinesin-5 function through control of neck-linker conformation. A kinesin-5 mutant lacking this N-terminus is enzymatically active, and ATP-dependent neck-linker movement and motility is defective although not ablated. All these aspects of kinesin-5 mechanochemistry are distinct from kinesin-1. Our findings directly demonstrate the regulatory role of the kinesin-5 N-terminus in collaboration with the motor’s structured neck-linker, and highlight the multiple adaptations within kinesin motor domains that tune their mechanochemistries according to distinct functional requirements.
Metadata
| Item Type: | Article |
|---|---|
| Keyword(s) / Subject(s): | cryo-electron microscopy, kinesins, mechanochemistry, microtubules, mitosis |
| School: | Birkbeck Faculties and Schools > Faculty of Science > School of Natural Sciences |
| Research Centres and Institutes: | Structural Molecular Biology, Institute of (ISMB) |
| Depositing User: | Administrator |
| Date Deposited: | 24 Jan 2014 12:30 |
| Last Modified: | 02 Aug 2023 17:08 |
| URI: | https://eprints.bbk.ac.uk/id/eprint/8898 |
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