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

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 pat...

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Main Authors: 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
Format: Article
Language:English
Published: eLife Sciences Publications Ltd 2025-05-01
Series:eLife
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Online Access:https://elifesciences.org/articles/105150
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author 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
author_facet 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
author_sort Elise Miedlar
collection DOAJ
description 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.
format Article
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issn 2050-084X
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spelling doaj-art-7ed477a8eebd48dca281955dbe5dfc732025-08-20T03:22:19ZengeLife Sciences Publications LtdeLife2050-084X2025-05-011410.7554/eLife.105150Force transmission through the inner kinetochore is enhanced by centromeric DNA sequencesElise Miedlar0https://orcid.org/0009-0009-8209-5210Grace E Hamilton1https://orcid.org/0000-0002-0522-0702Samuel R Witus2https://orcid.org/0000-0003-1907-8484Sara J Gonske3https://orcid.org/0009-0003-5491-9469Michael Riffle4https://orcid.org/0000-0003-1633-8607Alex Zelter5https://orcid.org/0000-0002-5331-0577Rachel E Klevit6https://orcid.org/0000-0002-3476-969XCharles L Asbury7https://orcid.org/0000-0002-0143-5394Yoana N Dimitrova8Trisha N Davis9https://orcid.org/0000-0003-4797-3152Department of Biochemistry, University of Washington, Seattle, United StatesDepartment of Biochemistry, University of Washington, Seattle, United StatesDepartment of Biochemistry, University of Washington, Seattle, United StatesDepartment of Biochemistry, University of Washington, Seattle, United StatesDepartment of Biochemistry, University of Washington, Seattle, United StatesDepartment of Biochemistry, University of Washington, Seattle, United StatesDepartment of Biochemistry, University of Washington, Seattle, United StatesDepartment of Neurobiology and Biophysics, University of Washington, Seattle, United StatesDepartment of Structural Biology, Genentech, Inc, South San Francisco, United StatesDepartment of Biochemistry, University of Washington, Seattle, United StatesPreviously, 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.https://elifesciences.org/articles/105150mitosiscentromerekinetochoremicrotubuletensionnucleosome
spellingShingle 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
Force transmission through the inner kinetochore is enhanced by centromeric DNA sequences
eLife
mitosis
centromere
kinetochore
microtubule
tension
nucleosome
title Force transmission through the inner kinetochore is enhanced by centromeric DNA sequences
title_full Force transmission through the inner kinetochore is enhanced by centromeric DNA sequences
title_fullStr Force transmission through the inner kinetochore is enhanced by centromeric DNA sequences
title_full_unstemmed Force transmission through the inner kinetochore is enhanced by centromeric DNA sequences
title_short Force transmission through the inner kinetochore is enhanced by centromeric DNA sequences
title_sort force transmission through the inner kinetochore is enhanced by centromeric dna sequences
topic mitosis
centromere
kinetochore
microtubule
tension
nucleosome
url https://elifesciences.org/articles/105150
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