Miedlar Elise, Hamilton Grace E, Witus Samuel R, Gonske Sara J, Riffle Michael, Zelter Alex, Klevit Rachel E, Asbury Charles L, Dimitrova Yoana N, Davis Trisha N
Department of Biochemistry, University of Washington, Seattle, United States.
Department of Neurobiology and Biophysics, University of Washington, Seattle, United States.
Elife. 2025 May 29;14:RP105150. doi: 10.7554/eLife.105150.
Previously, we reconstituted a minimal functional kinetochore from recombinant 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.
此前，我们利用重组蛋白重建了一个最小功能动粒，它能够将来自动态微管的力传递到含有着丝粒特异性组蛋白变体Cse4的核小体上（汉密尔顿等人，2020年）。这项工作揭示了通过内动粒进行力传递的两条途径：通过Mif2和通过Okp1/Ame1复合体（OA）。在这里，我们使用了一个嵌合DNA序列，它包含芽殖酵母点着丝粒的关键着丝粒决定元件，证明了含Cse4的核小体中着丝粒DNA序列的存在显著增强了OA介导的连接。我们的研究结果表明，着丝粒序列对于基于微管的力向染色体的传递很重要。
