1. Biochemistry and Chemical Biology
  2. Cell Biology
Download icon

Reconstitution reveals two paths of force transmission through the kinetochore

  1. Grace Elizabeth Hamilton
  2. Luke A Helgeson
  3. Cameron L Noland
  4. Charles L Asbury  Is a corresponding author
  5. Yoana N Dimitrova  Is a corresponding author
  6. Trisha N Davis  Is a corresponding author
  1. University of Washington, United States
  2. Genentech Inc, United States
Research Article
  • Cited 0
  • Views 375
  • Annotations
Cite this article as: eLife 2020;9:e56582 doi: 10.7554/eLife.56582

Abstract

Partitioning duplicated chromosomes equally between daughter cells is a microtubule-mediated process essential to eukaryotic life. A multi-protein machine, the kinetochore, drives chromosome segregation by coupling the chromosomes to dynamic microtubule tips, even as the tips grow and shrink through the gain and loss of subunits. The kinetochore must harness, transmit, and sense mitotic forces, as a lack of tension signals incorrect chromosome-microtubule attachment and precipitates error correction mechanisms. But though the field has arrived at a 'parts list' of dozens of kinetochore proteins organized into subcomplexes, the path of force transmission through these components has remained unclear. Here we report reconstitution of functional Saccharomyces cerevisiae kinetochore assemblies from recombinantly expressed proteins. The reconstituted kinetochores are capable of self-assembling in vitro, coupling centromeric nucleosomes to dynamic microtubules, and withstanding mitotically relevant forces. They reveal two distinct pathways of force transmission and Ndc80c recruitment.

Article and author information

Author details

  1. Grace Elizabeth Hamilton

    Department of Biochemistry, University of Washington, Seattle, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0522-0702
  2. Luke A Helgeson

    Department of Biochemistry, University of Washington, Seattle, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5112-2751
  3. Cameron L Noland

    Department of Structural Biology, Genentech Inc, South San Francisco, United States
    Competing interests
    Cameron L Noland, is affiliated with Genentech Inc. The author has no financial interests to declare.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6364-3167
  4. Charles L Asbury

    Department of Physiology and Biophysics, University of Washington, Seattle, United States
    For correspondence
    casbury@uw.edu
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0143-5394
  5. Yoana N Dimitrova

    Department of Structural Biology, Genentech Inc, South San Francisco, United States
    For correspondence
    dimitry4@gene.com
    Competing interests
    Yoana N Dimitrova, is affiliated with Genentech Inc. The author has no financial interests to declare.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1547-5781
  6. Trisha N Davis

    Department of Biochemistry, University of Washington, Seattle, United States
    For correspondence
    tdavis@uw.edu
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4797-3152

Funding

National Institutes of Health (Training Grant in Molecular Biophysics T32 GM008268)

  • Grace Elizabeth Hamilton

National Institutes of Health (R01 GM040506)

  • Trisha N Davis

National Institutes of Health (R35 GM130293)

  • Trisha N Davis

National Institutes of Health (R01 GM079373)

  • Charles L Asbury

National Institutes of Health (R35 GM134842)

  • Charles L Asbury

Genentech

  • Yoana N Dimitrova

Genentech

  • Cameron L Noland

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

Reviewing Editor

  1. Silke Hauf, Virginia Tech, United States

Publication history

  1. Received: March 3, 2020
  2. Accepted: May 13, 2020
  3. Accepted Manuscript published: May 14, 2020 (version 1)

Copyright

© 2020, Hamilton et al.

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.

Metrics

  • 375
    Page views
  • 126
    Downloads
  • 0
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Download citations (links to download the citations from this article in formats compatible with various reference manager tools)

Open citations (links to open the citations from this article in various online reference manager services)

Further reading

    1. Biochemistry and Chemical Biology
    2. Cell Biology
    Slavica Pavlovic Djuranovic et al.
    Research Article
    1. Biochemistry and Chemical Biology
    Santosh Kumar Kuncha et al.
    Research Article