Kinetochore-generated pushing forces separate centrosomes during bipolar spindle assembly

Abstract

In animal somatic cells, bipolar spindle formation requires separation of the centrosome-based spindle poles. Centrosome separation relies on multiple pathways, including cortical forces and antiparallel microtubule (MT) sliding, which are two activities controlled by the protein kinase aurora A. We previously found that depletion of the human kinetochore protein Mcm21R CENP-O results in monopolar spindles, raising the question as to whether kinetochores contribute to centrosome separation. In this study, we demonstrate that kinetochores promote centrosome separation after nuclear envelope breakdown by exerting a pushing force on the kinetochore fi bers (k-fi bers), which are bundles of MTs that connect kinetochores to centrosomes. This force is based on poleward MT fl ux, which incorporates new tubulin subunits at the plus ends of k-fi bers and requires stable k-fi bers to drive centrosomes apart. This kinetochore-dependent force becomes essential for centrosome separation if aurora A is inhibited. We conclude that two mechanisms control centrosome separation during prometaphase: an aurora A - dependent pathway and a kinetochore-dependent pathway that relies on k-fi ber-generated pushing forces. © 2009 Toso et al.