title: A biosensor of local kinesin activity reveals roles of PKC and EB1 in KIF17 activation.
abstract: We showed previously that the kinesin-2 motor KIF17 regulates microtubule (MT) dynamics and organization to promote epithelial differentiation. How KIF17 activity is regulated during this process remains unclear. Several kinesins, including KIF17, adopt compact and extended conformations that reflect autoinhibited and active states, respectively. We designed biosensors of KIF17 to monitor its activity directly in single cells using fluorescence lifetime imaging to detect Förster resonance energy transfer. Lifetime data are mapped on a phasor plot, allowing us to resolve populations of active and inactive motors in individual cells. Using this biosensor, we demonstrate that PKC contributes to the activation of KIF17 and that this is required for KIF17 to stabilize MTs in epithelia. Furthermore, we show that EB1 recruits KIF17 to dynamic MTs, enabling its accumulation at MT ends and thus promoting MT stabilization at discrete cellular domains.
authors.affiliation: Department of Cell and Developmental Biology, Weill Cornell Medical College of Cornell University, New York, NY 10021.
authors.forename: Cedric
authors.initials: C
authors.lastname: Espenel
authors.forename: Bipul R
authors.initials: BR
authors.lastname: Acharya
authors.forename: Geri
authors.initials: G
authors.lastname: Kreitzer
mesh_terms: D000818:Animals; D015374:Biosensing Techniques; D002460:Cell Line; D004285:Dogs; D004789:Enzyme Activation; D016547:Kinesins / Q000378:metabolism*; D061985:Madin Darby Canine Kidney Cells; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D011485:Protein Binding; D011493:Protein Kinase C / Q000378:metabolism*
chemical_list: C405804:EB1 microtubule binding proteins; D008869:Microtubule-Associated Proteins; D011493:Protein Kinase C; D016547:Kinesins