Visualisation of a kinesin-13 motor on microtubule end mimics

JOURNAL OF MOLECULAR BIOLOGY

卷:377

Article

Moores, Carolyn A.¹  Milligan, Ronald A.² 

[1] Univ London Birkbeck Coll, Sch Crystallog, London WC1E 7HX, England

[2] Scripps Res Inst, Dept Cell Biol, CB227, La Jolla, CA 92037 USA

关键词: depolymerisation;    dynamics;    electron microscopy;    kinesin-13;    microtubule;    tip-tracking proteins;   

DOI  :  10.1016/j.jmb.2008.01.079

来源: Elsevier

【 摘 要 】

An expanding collection of proteins localises to microtubule ends to regulate cytoskeletal dynamics and architecture by unknown molecular mechanisms. Electron microscopy is invaluable for studying microtubule structure, but because microtubule ends are heterogeneous, their structures are difficult to determine. We therefore investigated whether tubulin oligomers induced by the drug dolastatin could mimic microtubule ends. The microtubule end-dependent ATPase of kinesin-13 motors is coupled to microtubule depolymerisation. Significantly, kinesin-13 motor ATPase activity is stimulated by dolastatin-tubulin oligomers, suggesting, first, that these oligomers share properties with microtubule ends and, second, that the physical presence of an end is less important than terminal tubulin flexibility for microtubule end recognition by the kinesin-13 motor. Using electron microscopy, we visualised the kinesin-13 motor-dolastatin-tubulin oligomer interaction in nucleotide states mimicking steps in the ATPase cycle. This enabled us to detect conformational changes that the motor undergoes during depolymerisation. Our data suggest that such tubulin oligomers can be used to examine other microtubule end-binding proteins. (C) 2008 Elsevier Ltd. All rights reserved.

【 授权许可】

Free