The minus-end depolymerase KIF2A drives flux-like treadmilling of γTuRC-uncapped microtubules

Henkin, Brito, et al. reconstitute in vitro a functional module elucidating previously poorly understood mechanisms of poleward microtubule flux during eukaryotic cell division, reliant on the newly found inherent microtubule minus-end depolymerase activity of KIF2A, the major microtubule nucleator gamma TuRC, and a microtubule severing enzyme. During mitosis, microtubules in the spindle turn over continuously. At spindle poles, where microtubule minus ends are concentrated, microtubule nucleation and depolymerization, the latter required for poleward microtubule flux, happen side by side. How these seemingly antagonistic processes of nucleation and depolymerization are coordinated is not understood. Here, we reconstitute this coordination in vitro combining different pole-localized activities. We find that the spindle pole-localized kinesin-13 KIF2A is a microtubule minus-end depolymerase, in contrast to its paralog MCAK. Due to its asymmetric activity, KIF2A still allows microtubule nucleation from the gamma-tubulin ring complex (gamma TuRC), which serves as a protective cap shielding the minus end against KIF2A binding. Efficient gamma TuRC uncapping requires the combined action of KIF2A and a microtubule severing enzyme, leading to treadmilling of the uncapped microtubule driven by KIF2A. Together, these results provide insight into the molecular mechanisms by which a minimal protein module coordinates microtubule nucleation and depolymerization at spindle poles consistent with their role in poleward microtubule flux.

Automated summary

Researchers reconstitute a functional module in vitro, shedding light on previously poorly understood mechanisms of poleward microtubule flux during eukaryotic cell division. This mechanism relies on the newly discovered inherent microtubule minus-end depolymerase activity of KIF2A, the major microtubule nucleator gamma TuRC, and a microtubule severing enzyme.