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.
authors.affiliation: Department of Cell and Developmental Biology, University of Michigan, 109 Zina Pitcher Place, Ann Arbor, Michigan 48109, USA.
authors.forename: Martin F
authors.initials: MF
authors.lastname: Engelke
authors.affiliation: Department of Cell and Molecular Biology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois 60611, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Winding
authors.affiliation: Department of Cell and Developmental Biology, University of Michigan, 109 Zina Pitcher Place, Ann Arbor, Michigan 48109, USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Yue
authors.affiliation: Department of Biomedical Engineering, Penn State University, University Park, Pennsylvania 16802, USA.
authors.forename: Shankar
authors.initials: S
authors.lastname: Shastry
authors.affiliation: Department of Cell and Developmental Biology, University of Michigan, 109 Zina Pitcher Place, Ann Arbor, Michigan 48109, USA.
authors.forename: Federico
authors.identifier: http://orcid.org/0000-0003-0953-8010
authors.initials: F
authors.lastname: Teloni
authors.affiliation: Department of Cell and Developmental Biology, University of Michigan, 109 Zina Pitcher Place, Ann Arbor, Michigan 48109, USA.
authors.forename: Sanjay
authors.initials: S
authors.lastname: Reddy
authors.affiliation: Department of Cell and Developmental Biology, University of Michigan, 109 Zina Pitcher Place, Ann Arbor, Michigan 48109, USA.
authors.forename: T Lynne
authors.initials: TL
authors.lastname: Blasius
authors.affiliation: Department of Cell and Developmental Biology, University of Michigan, 109 Zina Pitcher Place, Ann Arbor, Michigan 48109, USA.
authors.forename: Pushpanjali
authors.initials: P
authors.lastname: Soppina
authors.affiliation: Department of Biomedical Engineering, Penn State University, University Park, Pennsylvania 16802, USA.
authors.forename: William O
authors.initials: WO
authors.lastname: Hancock
authors.affiliation: Department of Cell and Molecular Biology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois 60611, USA.
authors.forename: Vladimir I
authors.initials: VI
authors.lastname: Gelfand
authors.affiliation: Department of Cell and Developmental Biology, University of Michigan, 109 Zina Pitcher Place, Ann Arbor, Michigan 48109, USA.
authors.forename: Kristen J
authors.initials: KJ
authors.lastname: Verhey
mesh_terms: D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D002522:Chlorocebus aethiops; D004331:Drosophila melanogaster; D004398:Dyneins / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D016547:Kinesins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008870:Microtubules / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D057054:Molecular Imaging; D009218:Myosins / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D015202:Protein Engineering; D021381:Protein Transport; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D054852:Small Molecule Libraries / Q000138:chemical synthesis / Q000494:pharmacology*; D014162:Transfection; D050257:Tubulin Modulators / Q000138:chemical synthesis / Q000494:pharmacology*
chemical_list: C117477:KIF25 protein, human; C488101:KIF5B protein, human; D011994:Recombinant Proteins; D054852:Small Molecule Libraries; D050257:Tubulin Modulators; D009218:Myosins; D004398:Dyneins; D016547:Kinesins