Interplay of Microtubule Dynamics and Sliding during Bipolar Spindle Formation in Mammalian Cells

Accurate chromosome segregation during mitosis relies on the organization of microtubules into a bipolar spindle. Kinesin-5 proteins play an evolutionarily conserved role in establishing spindle bipolarity [1, 2] and clinical trials are currently evaluating inhibitors of human kinesin-5 (i.e., Eg5) for chemotherapeutic potential. However, in mammalian somatic cells, Eg5 activity is dispensable for maintenance of bipolar spindles once they are formed [3, 4], suggesting distinct requirements for establishment versus maintenance of spindle bipolarity. By combining Eg5 inhibition with RNA interference of other spindle proteins, we show that mitotic cells deficient in MCAK fail to maintain spindle bipolarity in the absence of Eg5 activity. Collapse of bipolar spindles in MCAK-deficient cells is driven by pole-focusing activities and is independent of MCAK function at centromeres, implicating hyperstabilized non-kinetochore microtubules in spindle collapse. Conversely, destabilizing nonkinetochore microtubules in early mitosis reduces the reliance on Eg5 for establishment of spindle bipolarity and renders cells partially resistant to Eg5 inhibitors. Thus, the temporal requirement for microtubule sliding generated by Eg5 activity during bipolar spindle assembly in mammalian cells is regulated by changes in the dynamic behavior of microtubules during mitosis.