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A quantitative description of Ndc80 complex linkage to human kinetochores

Author

Listed:
  • Aussie Suzuki

    (University of North Carolina at Chapel Hill)

  • Benjamin L. Badger

    (University of North Carolina at Chapel Hill)

  • Edward D. Salmon

    (University of North Carolina at Chapel Hill)

Abstract

The Ndc80 complex, which mediates end-on attachment of spindle microtubules, is linked to centromeric chromatin in human cells by two inner kinetochore proteins, CENP-T and CENP-C. Here to quantify their relative contributions to Ndc80 recruitment, we combine measurements of kinetochore protein copy number with selective protein depletion assays. This approach reveals about 244 Ndc80 complexes per human kinetochore (∼14 per kinetochore microtubule), 215 CENP-C, 72 CENP-T and only 151 Ndc80s as part of the KMN protein network (1:1:1 Knl1, Mis12 and Ndc80 complexes). Each CENP-T molecule recruits ∼2 Ndc80 complexes; one as part of a KMN network. In contrast, ∼40% of CENP-C recruits only a KMN network. Replacing the CENP-C domain that binds KMN with the CENP-T domain that recruits both an Ndc80 complex and KMN network yielded functional kinetochores. These results provide a quantitative picture of the linkages between centromeric chromatin and the microtubule-binding Ndc80 complex at the human kinetochore.

Suggested Citation

  • 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.
    • Handle: RePEc:nat:natcom:v:6:y:2015:i:1:d:10.1038_ncomms9161
    DOI: 10.1038/ncomms9161
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    Cited by:

    1. 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.

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