Rules of engagement: centrosome-nuclear connections in a closed mitotic system.

The assembly of a functional mitotic spindle is essential for cell reproduction and requires a precise coordination between the nuclear cycle and the centrosome. This coordination is particularly prominent in organisms that undergo closed mitosis where centrosomes must not only respond to temporal signals, but also to spatial considerations, e.g. switching from the production of cytoplasmic microtubule arrays to the generation of dynamic intra-nuclear microtubules required for spindle assembly. We utilize a gene knockout of Kif9, a Dictyostelium discoideum Kin-I kinesin, to destabilize the physical association between centrosomes and the nuclear envelope. This approach presents a unique opportunity to reveal temporal and spatial components in the regulation of centrosomal activities in a closed-mitosis organism. Here we report that centrosome-nuclear engagement is not required for the entry into mitosis. Although detached centrosomes can duplicate in the cytoplasm, neither they nor nuclei alone can produce spindle-like microtubule arrays. However, the physical association of centrosomes and the nuclear envelope is required to progress through mitosis beyond prometaphase.

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