A structural model for microtubule minus-end recognition and protection by CAMSAP proteins
Atherton, Joseph; Jiang, Kai; Stangier, Marcel M.; Luo, Yanzhang; Hua, Shasha; Houben, Klaartje; Van Hooff, Jolien J.E.; Joseph, Agnel Praveen; Scarabelli, Guido; Grant, Barry J.; Roberts, Anthony J.; Topf, Maya; Steinmetz, Michel O.; Baldus, Marc; Moores, Carolyn A.; Akhmanova, Anna
(2017) Nature Structural and Molecular Biology, volume 24, issue 11, pp. 931 - 943
(Article)
Abstract
CAMSAP and Patronin family members regulate microtubule minus-end stability and localization and thus organize noncentrosomal microtubule networks, which are essential for cell division, polarization and differentiation. Here, we found that the CAMSAP C-terminal CKK domain is widely present among eukaryotes and autonomously recognizes microtubule minus ends. Through a combination of
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structural approaches, we uncovered how mammalian CKK binds between two tubulin dimers at the interprotofilament interface on the outer microtubule surface. In vitro reconstitution assays combined with high-resolution fluorescence microscopy and cryo-electron tomography suggested that CKK preferentially associates with the transition zone between curved protofilaments and the regular microtubule lattice. We propose that minus-end-specific features of the interprotofilament interface at this site serve as the basis for CKK's minus-end preference. The steric clash between microtubule-bound CKK and kinesin motors explains how CKK protects microtubule minus ends against kinesin-13-induced depolymerization and thus controls the stability of free microtubule minus ends.
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Keywords: Cryoelectron microscopy, Cryoelectron tomography, Kinesin, Microtubules, NMR spectroscopy, Structural Biology, Molecular Biology
ISSN: 1545-9993
Publisher: Nature Publishing Group
Note: Funding Information: We thank F. Govers (Wageningen University), V. Gelfand (Northwestern University) and M. Harterink (Utrecht University) for sharing reagents; E. Katrukha for advice on processing fluorescence microscopy data; R. Boelens for providing access to the solution-state NMR instrumentation; and S. Yokoyama and A. Nomura for sharing NMR resonance assignments of free CKK. A.A. was supported by ERC Synergy grant 609822. J.A., C.A.M. (MR/J000973/1), A.-P.J. and M.T. (MR/M019292/1) were supported by the Medical Research Council, UK. A.J.R. was supported by a Sir Henry Dale Fellowship from the Wellcome Trust and Royal Society (104196/Z/14/Z). G.S. and B.J.G. were supported by the National Institutes of Health (R01GM070862). M.B., Y.L. (grant 718.015.001) and K.H. (grant 718.015.001 and 184.032.207) were supported by the Netherlands Organization for Scientific Research (NWO). ssNMR experiments were supported by uNMR-NL, an NWO-funded National Roadmap Large-scale Facility of the Netherlands (grant 184.032.207). M.O.S. was supported by grants from the Swiss National Science Foundation (31003A_ 166608) and from SystemsX.ch (RTD-TubeX). Publisher Copyright: © 2017 Nature America, Inc., part of Springer Nature. All rights reserved.
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