Force transmission through the inner kinetochore is enhanced by centromeric DNA sequences
2025
Elise Miedlar | Grace E Hamilton | Samuel R Witus | Sara J Gonske | Michael Riffle | Alex Zelter | Rachel E Klevit | Charles L Asbury | Yoana N Dimitrova | Trisha N Davis
Previously, we reconstituted a minimal functional kinetochore from recombinant Saccharomyces 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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