Engineered kinesin motor proteins amenable to small-molecule inhibition

Martin F. Engelke; Michael Winding; Yang Yue; Shankar Shastry; Federico Teloni; Sanjay Reddy; T. Lynne Blasius; Pushpanjali Soppina; William O. Hancock; Vladimir I. Gelfand; Kristen J. Verhey

doi:10.1038/ncomms11159

Engineered kinesin motor proteins amenable to small-molecule inhibition

Martin F. Engelke, Michael Winding, Yang Yue, Shankar Shastry, Federico Teloni, Sanjay Reddy, T. Lynne Blasius, Pushpanjali Soppina, William O. Hancock, Vladimir I. Gelfand, Kristen J. Verhey

Research output: Contribution to journal › Article › peer-review

25 Scopus citations

Abstract

The human genome encodes 45 kinesin motor proteins that drive cell division, cell motility, intracellular trafficking and ciliary function. Determining the cellular function of each kinesin would benefit from specific small-molecule inhibitors. However, screens have yielded only a few specific inhibitors. Here we present a novel chemical-genetic approach to engineer kinesin motors that can carry out the function of the wild-type motor yet can also be efficiently inhibited by small, cell-permeable molecules. Using kinesin-1 as a prototype, we develop two independent strategies to generate inhibitable motors, and characterize the resulting inhibition in single-molecule assays and in cells. We further apply these two strategies to create analogously inhibitable kinesin-3 motors. These inhibitable motors will be of great utility to study the functions of specific kinesins in a dynamic manner in cells and animals. Furthermore, these strategies can be used to generate inhibitable versions of any motor protein of interest.

Original language	English (US)
Article number	11159
Journal	Nature communications
Volume	7
DOIs	https://doi.org/10.1038/ncomms11159
State	Published - Apr 5 2016

All Science Journal Classification (ASJC) codes

General Chemistry
General Biochemistry, Genetics and Molecular Biology
General Physics and Astronomy

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10.1038/ncomms11159

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title = "Engineered kinesin motor proteins amenable to small-molecule inhibition",

abstract = "The human genome encodes 45 kinesin motor proteins that drive cell division, cell motility, intracellular trafficking and ciliary function. Determining the cellular function of each kinesin would benefit from specific small-molecule inhibitors. However, screens have yielded only a few specific inhibitors. Here we present a novel chemical-genetic approach to engineer kinesin motors that can carry out the function of the wild-type motor yet can also be efficiently inhibited by small, cell-permeable molecules. Using kinesin-1 as a prototype, we develop two independent strategies to generate inhibitable motors, and characterize the resulting inhibition in single-molecule assays and in cells. We further apply these two strategies to create analogously inhibitable kinesin-3 motors. These inhibitable motors will be of great utility to study the functions of specific kinesins in a dynamic manner in cells and animals. Furthermore, these strategies can be used to generate inhibitable versions of any motor protein of interest.",

author = "Engelke, {Martin F.} and Michael Winding and Yang Yue and Shankar Shastry and Federico Teloni and Sanjay Reddy and Blasius, {T. Lynne} and Pushpanjali Soppina and Hancock, {William O.} and Gelfand, {Vladimir I.} and Verhey, {Kristen J.}",

note = "Funding Information: Acknowledgements We thank S. Norris, V. Soppina, D. Cai and D. Nunez (University of Michigan) for support with TIRF imaging and/or data analysis and C. Hoogenraad (Utrecht University) and M. Rolls (Penn State University) for sharing plasmids. This work was supported by postdoc mobility fellowships (PBZHP3-141433 and P300P3-154631) from the Swiss National Science Foundation to M.F.E. and by NIH grants R21NS078761 and R01GM070862 to K.J.V. and R01GM52112 to V.I.G.",

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AU - Reddy, Sanjay

AU - Blasius, T. Lynne

AU - Soppina, Pushpanjali

AU - Hancock, William O.

AU - Gelfand, Vladimir I.

AU - Verhey, Kristen J.

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N2 - The human genome encodes 45 kinesin motor proteins that drive cell division, cell motility, intracellular trafficking and ciliary function. Determining the cellular function of each kinesin would benefit from specific small-molecule inhibitors. However, screens have yielded only a few specific inhibitors. Here we present a novel chemical-genetic approach to engineer kinesin motors that can carry out the function of the wild-type motor yet can also be efficiently inhibited by small, cell-permeable molecules. Using kinesin-1 as a prototype, we develop two independent strategies to generate inhibitable motors, and characterize the resulting inhibition in single-molecule assays and in cells. We further apply these two strategies to create analogously inhibitable kinesin-3 motors. These inhibitable motors will be of great utility to study the functions of specific kinesins in a dynamic manner in cells and animals. Furthermore, these strategies can be used to generate inhibitable versions of any motor protein of interest.

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Engineered kinesin motor proteins amenable to small-molecule inhibition

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