NuMA is a mitotic adaptor protein that activates dynein and connects it to microtubule minus ends

Abstract

ABSTRACTNuclear mitotic apparatus protein (NuMA) is indispensable for the mitotic functions of the major microtubule minus-end directed motor cytoplasmic dynein 1. NuMA and dynein are both essential for correct spindle pole organization. How these proteins cooperate to gather microtubule minus ends at spindle poles remains unclear. Here we use microscopy-basedin vitroreconstitutions to demonstrate that NuMA is a dynein adaptor, activating processive dynein motility together with dynein’s cofactors dynactin and Lissencephaly-1 (Lis1). Additionally, we find that NuMA binds and stabilizes microtubule minus ends, allowing dynein/dynactin/NuMA to transport microtubule minus ends as cargo to other minus ends. We further show that the microtubule-nucleating γ-tubulin ring complex (γTuRC) hinders NuMA binding. This shows that either γTuRC needs to be released or microtubules need to be severed to generate free minus ends before they can be incorporated into spindle poles by dynein/dynactin/NuMA. These results provide new mechanistic insight into how dynein, dynactin, NuMA and Lis1 cooperate to organize spindle poles in cells.