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April 1st, 2025
Version: 3
University of Washington
biochemistry
biorxiv

Force transmission through the inner kinetochore is enhanced by centromeric DNA sequences

Miedlar, E.Open in Google Scholar•Hamilton, G. E.Open in Google Scholar•Witus, S. R.Open in Google Scholar•Gonske, S. J.Open in Google Scholar•Riffle, M.Open in Google Scholar•Zelter, A.Open in Google Scholar•Klevit, R. E.Open in Google Scholar•Asbury, C. L.Open in Google Scholar•Dimitrova, Y. N.Open in Google Scholar•Davis, T. N.Open in Google Scholar

Previously, we reconstituted a minimal functional kinetochore from recombinant S. cerevisiae proteins that was capable of transmitting force from dynamic microtubules to nucleosomes containing the centromere-specific histone variant Cse4 (Hamilton et al. 2020). This work revealed two paths of force transmission through the inner kinetochore: through Mif2 and through the Okp1/Ame1 complex (OA). Here, using a chimeric DNA sequence that contains crucial centromere-determining elements of the budding yeast point centromere, we demonstrate that the presence of centromeric DNA sequences in Cse4-containing nucleosomes significantly strengthens OA-mediated linkages. Our findings indicate that centromeric sequences are important for the transmission of microtubule-based forces to the chromosome.

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