Abstract
Coupling between ATPase and track binding sites is essential for molecular motors to move along cytoskeletal tracks. In dynein, these sites are separated by a long coiled coil stalk that must mediate communication between them, but the underlying mechanism remains unclear. Here we show that changes in registration between the two helices of the coiled coil can perform this function. We locked the coiled coil at three specific registrations using oxidation to disulfides of paired cysteine residues introduced into the two helices. These trapped ATPase activity either in a microtubule-independent high or low state, and microtubule binding activity either in an ATP-insensitive strong or weak state, depending on the registry of the coiled coil. Our results provide direct evidence that dynein uses sliding between the two helices of the stalk to couple ATPase and microtubule binding activities during its mechanochemical cycle.
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Acknowledgements
We thank M. Koonce (Wadsworth Center) for providing the anti-stalk antibody. This work was supported by a Grant-in-Aid for Scientific Research (S) from the Japan Society for the Promotion of Science (JSPS) to K.S., a Grant-in-Aid for Scientific Research on Priority Areas from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (MEXT) to K.S., a Grant-in-Aid for Young Scientists (A) from MEXT to T.K., and in part by The Wellcome Trust to A.J.R., the Biotechnology and Biological Sciences Research Council (UK) to S.A.B. and P.J.K., and Grant GM30401 from the US National Institutes of Health to I.R.G.
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T.K., I.R.G and K.S. designed the study; T.K., K.I. and R.O. performed the biochemical experiments; A.J.R., P.J.K. and S.A.B. performed the EM analyses; K.S. supervised the study; T.K. wrote the first draft of the manuscript and all authors contributed to the preparation of the final version.
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Kon, T., Imamula, K., Roberts, A. et al. Helix sliding in the stalk coiled coil of dynein couples ATPase and microtubule binding. Nat Struct Mol Biol 16, 325–333 (2009). https://doi.org/10.1038/nsmb.1555
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DOI: https://doi.org/10.1038/nsmb.1555
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