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Significance Eg5, a mitotic kinesin, has long been an anticancer drug target. However, a different kinesin motor, KIF15, can rescue cell division when Eg5 is incapacitated, leading to chemotherapeutic resistance. We characterized KIF15 motor mechanics at the single-molecule level and studied the effects of combinations of small-molecule inhibitors of KIF15 and Eg5 on admixtures of motors in a motility assay, as well as on cancer cell proliferation. Taken together, our results point the way toward a strategy of combination drug therapy targeting both Eg5 and KIF15 as a means of overcoming KIF15-mediated cancer resistance. This work highlights the importance of understanding the molecular physiology of different kinesins and of exploring inhibitors that target additional mitotic kinesins.

KIF15 nanomechanics and kinesin inhibitors, with implications for cancer chemotherapeutics