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CENP-F controls force generation and the dynein-dependent stripping of CENP-E at kinetochores

View ORCID ProfilePhilip Auckland, View ORCID ProfileAndrew D. McAinsh
doi: https://doi.org/10.1101/627380
Philip Auckland
Centre for Mechanochemical Cell Biology & Division of Biomedical Sciences, Warwick Medical School, The University of Warwick, Gibbet Hill Road, Coventry, CV4 7AL, UK.
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  • For correspondence: a.d.mcainsh@warwick.ac.uk p.auckland.1@warwick.ac.uk
Andrew D. McAinsh
Centre for Mechanochemical Cell Biology & Division of Biomedical Sciences, Warwick Medical School, The University of Warwick, Gibbet Hill Road, Coventry, CV4 7AL, UK.
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  • For correspondence: a.d.mcainsh@warwick.ac.uk p.auckland.1@warwick.ac.uk
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Abstract

Accurate chromosome segregation demands efficient capture of microtubules by kinetochores and their conversion to stable bi-oriented attachments that can congress and then segregate chromosomes. An early event is the shedding of the outermost fibrous corona layer of the kinetochore following microtubule attachment. Centromere protein F (CENP-F) is part of the corona, contains two microtubule-binding domains and physically associates with dynein motor regulators. Here, we have combined CRISPR gene editing and engineered separation-of-function mutants to define how CENP-F contributes to kinetochore function. We show here that the two microtubule-binding domains make distinct contributions to attachment stability and force generation that are required to minimise errors in anaphase, but are dispensable for congression. We further identify a specialised domain that functions to inhibit the dynein mediated stripping of CENP-E motors. We show how this “dynein-brake” is crucial for ensuring kinetochores contain the right number of CENP-E motors at the right time during mitosis, with loss of brake function delaying congression.

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Posted May 03, 2019.
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CENP-F controls force generation and the dynein-dependent stripping of CENP-E at kinetochores
Philip Auckland, Andrew D. McAinsh
bioRxiv 627380; doi: https://doi.org/10.1101/627380
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CENP-F controls force generation and the dynein-dependent stripping of CENP-E at kinetochores
Philip Auckland, Andrew D. McAinsh
bioRxiv 627380; doi: https://doi.org/10.1101/627380

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