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Recruitment of two Ndc80 complexes via the CENP-T pathway is sufficient for kinetochore functions

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  • Yusuke Takenoshita

    (Osaka University)

  • Masatoshi Hara

    (Osaka University)

  • Tatsuo Fukagawa

    (Osaka University)

Abstract

To form functional kinetochores, CENP-C and CENP-T independently recruit the KMN (Knl1C, Mis12C, and Ndc80C) network onto the kinetochores. To clarify the functions of the KMN network on CENP-T, we evaluated its roles in chicken DT40 cell lines lacking the CENP-C-KMN network interaction. By analyzing mutants lacking both CENP-T-Mis12C and CENP-C-Mis12C interactions, we demonstrated that Knl1C and Mis12C (KM) play critical roles in the cohesion of sister chromatids or the recruitment of spindle checkpoint proteins onto kinetochores. Two copies of Ndc80C (N-N) exist on CENP-T via Mis12C or direct binding. Analyses of cells specifically lacking the Mis12C-Ndc80C interaction revealed that N-N is needed for proper kinetochore-microtubule interactions. However, using artificial engineering to directly bind the two copies of Ndc80C to CENP-T, we demonstrated that N-N functions without direct Mis12C binding to Ndc80C in native kinetochores. This study demonstrated the mechanisms by which complicated networks play roles in native kinetochores.

Suggested Citation

  • Yusuke Takenoshita & Masatoshi Hara & Tatsuo Fukagawa, 2022. "Recruitment of two Ndc80 complexes via the CENP-T pathway is sufficient for kinetochore functions," Nature Communications, Nature, vol. 13(1), pages 1-19, December.
    • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-28403-8
    DOI: 10.1038/s41467-022-28403-8
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    References listed on IDEAS

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    1. Anna A. Ye & Stuart Cane & Thomas J. Maresca, 2016. "Chromosome biorientation produces hundreds of piconewtons at a metazoan kinetochore," Nature Communications, Nature, vol. 7(1), pages 1-9, December.
    2. Aussie Suzuki & Benjamin L. Badger & Edward D. Salmon, 2015. "A quantitative description of Ndc80 complex linkage to human kinetochores," Nature Communications, Nature, vol. 6(1), pages 1-14, November.
    3. Gregory M. Alushin & Vincent H. Ramey & Sebastiano Pasqualato & David A. Ball & Nikolaus Grigorieff & Andrea Musacchio & Eva Nogales, 2010. "The Ndc80 kinetochore complex forms oligomeric arrays along microtubules," Nature, Nature, vol. 467(7317), pages 805-810, October.
    4. Shreyas Sridhar & Tetsuya Hori & Reiko Nakagawa & Tatsuo Fukagawa & Kaustuv Sanyal, 2021. "Bridgin connects the outer kinetochore to centromeric chromatin," Nature Communications, Nature, vol. 12(1), pages 1-19, December.
    5. John R. Weir & Alex C. Faesen & Kerstin Klare & Arsen Petrovic & Federica Basilico & Josef Fischböck & Satyakrishna Pentakota & Jenny Keller & Marion E. Pesenti & Dongqing Pan & Doro Vogt & Sabine Woh, 2016. "Insights from biochemical reconstitution into the architecture of human kinetochores," Nature, Nature, vol. 537(7619), pages 249-253, September.
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