@article {Kern151399, author = {David M. Kern and Elizabeth M. Wilson-Kubalek and Iain M. Cheeseman}, title = {Astrin-SKAP complex reconstitution reveals its kinetochore interaction with microtubule-bound Ndc80}, elocation-id = {151399}, year = {2017}, doi = {10.1101/151399}, publisher = {Cold Spring Harbor Laboratory}, abstract = {Chromosome segregation requires robust interactions between the macromolecular kinetochore structure and dynamic microtubule polymers. A key outstanding question is how kinetochore-microtubule attachments are modulated to ensure that bi-oriented attachments are selectively stabilized and maintained. The Astrin-SKAP complex localizes preferentially to properly bi-oriented sister kinetochores, representing the final outer kinetochore component recruited prior to anaphase onset. Here, we reconstitute the 4-subunit Astrin-SKAP complex, including a novel MYCBP subunit. Our work demonstrates that the Astrin-SKAP complex contains separable kinetochore localization and microtubule binding domains. In addition, through cross-linking analysis in human cells and biochemical reconstitution, we show that the Astrin-SKAP complex binds synergistically to microtubules with the Ndc80 complex, the core component of the kinetochore-microtubule interface, to form an integrated interface. We propose a model in which the Astrin-SKAP complex acts together with the Ndc80 complex to stabilize correctly formed kinetochore-microtubule interactions.}, URL = {https://www.biorxiv.org/content/early/2017/06/17/151399}, eprint = {https://www.biorxiv.org/content/early/2017/06/17/151399.full.pdf}, journal = {bioRxiv} }