Mitotic spindle formation relies on the stochastic capture of microtubules at kinetochores. Kinetochore architecture affects the efficiency and fidelity of this process with large kinetochores expected to accelerate assembly at the expense of accuracy, and smaller kinetochores to suppress errors at the expense of efficiency. We demonstrate that upon mitotic entry, kinetochores in cultured human cells form large crescents that subsequently compact into discrete structures on opposite sides of the centromere. This compaction occurs only after the formation of end-on microtubule attachments. Live-cell microscopy reveals that centromere rotation mediated by lateral kinetochore-microtubule interactions precedes formation of end-on attachments and kinetochore compaction. Computational analyses of kinetochore expansion-compaction in the context of lateral interactions correctly predict experimentally-observed spindle assembly times with reasonable error rates. The computational model suggests that larger kinetochores reduce both errors and assembly times, which can explain the robustness of spindle assembly and the functional significance of enlarged kinetochores.

In the absence of continuous K-fiber attachment between each kinetochore and the spindle pole, one or more additional mechanisms dependent on dynein-mediated kinetochore transport exist to ensure proper chromosome segregation during mitosis.

Taxol induces extensive structural reorganization of the mammalian kinetochore; however, this reorganization is not sufficient to maintain a long-term mitotic arrest unless some of the kinetochores completely lose their attachment to microtubules.