The mitotic kinesin-14 KlpA contains a context-dependent directionality switch
Andrew R. Popchock, Kuo-Fu Tseng, Pan Wang, P. Andrew Karplus, Xin Xiang & Weihong Qiu
Nature Communications 8, Article number: 13999 (2017)
doi: 10.1038/ncomms13999
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KinesinSingle-molecule biophysics
Received: 16 June 2016 Accepted: 17 November 2016 Published online: 04 January 2017
Source/Fonte: Dynamic Science
Abstract
Kinesin-14s are commonly known as nonprocessive minus end-directed microtubule motors that function mainly for mitotic spindle assembly. Here we show using total internal reflection fluorescence microscopy that KlpA—a kinesin-14 from Aspergillus nidulans—is a context-dependent bidirectional motor. KlpA exhibits plus end-directed processive motility on single microtubules, but reverts to canonical minus end-directed motility when anchored on the surface in microtubule-gliding experiments or interacting with a pair of microtubules in microtubule-sliding experiments. Plus end-directed processive motility of KlpA on single microtubules depends on its N-terminal nonmotor microtubule-binding tail, as KlpA without the tail is nonprocessive and minus end-directed. We suggest that the tail is a de facto directionality switch for KlpA motility: when the tail binds to the same microtubule as the motor domain, KlpA is a plus end-directed processive motor; in contrast, when the tail detaches from the microtubule to which the motor domain binds, KlpA becomes minus end-directed.
Acknowledgements
We thank Drs C. Mathews (Oregon State University), X. Su (UCSF) and B. Liu (UC Davis) for critical reading of the manuscript, and Mr Chun Liu (Pearl River Fisheries Research Institute, China) for initial plasmid construction. This work was supported in part by the National Science Foundation (MCB-1616462 to W.Q.).
Author information
Author notes
Andrew R. Popchock & Kuo-Fu Tseng
These authors contributed equally to this work
Affiliations
Department of Biochemistry and Biophysics, Oregon State University, Corvallis, Oregon 97331, USA
Andrew R. Popchock, P. Andrew Karplus & Weihong Qiu
Department of Physics, Oregon State University, Corvallis, Oregon 97331, USA
Kuo-Fu Tseng, Pan Wang & Weihong Qiu
School of Physics and Electronics, Henan University, Kaifeng, Henan 475004, China
Pan Wang
Department of Biochemistry and Molecular Biology, The Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814, USA
Xin Xiang
Contributions
W.Q. conceived, designed and supervised the study; A.R.P. and K.-F.T. performed the experiments; K.-F.T. and P.W. contributed all KlpA constructs. All authors participated in discussing and interpreting the results. P.A.K., X.X. and W.Q. wrote the manuscript with input from all other authors.
Competing interests
The authors declare no competing financial interests.
Corresponding author
Correspondence to Weihong Qiu.
