Abstract

Eukaryotic kinetochores connect spindle microtubules to chromosomal centromeres. A group of proteins called the Ctf19 complex (Ctf19c) in yeast and the constitutive centromere associated network (CCAN) in other organisms creates the foundation of a kinetochore. The Ctf19c/CCAN influences the timing of kinetochore assembly, sets its location by associating with a specialized nucleosome containing the histone H3 variant Cse4/CENP-A, and determines the organization of the microtubule attachment apparatus. We present here the structure of a reconstituted 13-subunit Ctf19c determined by cryo-electron microscopy at ~4 Å resolution. The structure accounts for known and inferred contacts with the Cse4 nucleosome and for an observed assembly hierarchy. We describe its implications for establishment of kinetochores and for their regulation by kinases throughout the cell cycle.

Data availability

We have deposited the model coordinates and cryo-EM maps in the PDB (6NUW) and EMDB (EMD-0523). Tracking files for imaging experiments are included as a source data file associated with Figure 3.

The following data sets were generated

Article and author information

Author details

  1. Stephen M Hinshaw

    Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, United States
    For correspondence
    hinshaw@crystal.harvard.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4215-5206
  2. Stephen C Harrison

    Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, United States
    For correspondence
    harrison@crystal.harvard.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7215-9393

Funding

Howard Hughes Medical Institute

  • Stephen M Hinshaw
  • Stephen C Harrison

Helen Hay Whitney Foundation

  • Stephen M Hinshaw

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Andrea Musacchio, Max Planck Institute of Molecular Physiology, Germany

Version history

  1. Received: December 8, 2018
  2. Accepted: February 13, 2019
  3. Accepted Manuscript published: February 14, 2019 (version 1)
  4. Version of Record published: March 8, 2019 (version 2)
  5. Version of Record updated: March 5, 2020 (version 3)

Copyright

© 2019, Hinshaw & Harrison

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Stephen M Hinshaw
  2. Stephen C Harrison
(2019)
The structure of the Ctf19c/CCAN from budding yeast
eLife 8:e44239.
https://doi.org/10.7554/eLife.44239

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https://doi.org/10.7554/eLife.44239

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