The COMA complex interacts with Cse4 and positions Sli15/Ipl1 at the budding yeast inner kinetochore

  1. Josef Fischböck-Halwachs
  2. Sylvia Singh
  3. Mia Potocnjak
  4. Götz Hagemann
  5. Victor Solis-Mezarino
  6. Stephan Woike
  7. Medini Ghodgaonkar Steger
  8. Florian Weissmann
  9. Laura D Gallego
  10. Julie Rojas
  11. Jessica Andreani-Feuillet
  12. Alwin Köhler
  13. Franz Herzog  Is a corresponding author
  1. Ludwig-Maximilians-Universität München, Germany
  2. Vienna Biocenter, Germany
  3. Medical University of Vienna, Austria
  4. Max Planck Institute of Biochemistry, Germany
  5. University Paris-Sud, France

Abstract

Kinetochores are macromolecular protein complexes at centromeres that ensure accurate chromosome segregation by attaching chromosomes to spindle microtubules and integrating safeguard mechanisms. The inner kinetochore is assembled on CENP-A nucleosomes and has been implicated in establishing a kinetochore-associated pool of Aurora B kinase, a chromosomal passenger complex (CPC) subunit, which is essential for chromosome biorientation. By performing crosslink-guided in vitro reconstitution of budding yeast kinetochore complexes we showed that the Ame1/Okp1CENP-U/Q heterodimer, which forms the COMA complex with Ctf19/Mcm21CENP-P/O, selectively bound Cse4CENP-A nucleosomes through the Cse4 N-terminus. The Sli15/Ipl1INCENP/Aurora-B core-CPC interacted with COMA in vitro through the Ctf19 C-terminus whose deletion affects accurate chromosome segregation in a Sli15 wild-type background. Tethering Sli15 to Ame1/Okp1 rescued synthetic lethality upon Ctf19 depletion in a Sli15 centromere-targeting deficient mutant. This study shows molecular characteristics of the point-centromere inner kinetochore architecture and suggests a role for the Ctf19 C-terminus in mediating accurate chromosome segregation.

Data availability

The mass spectrometry raw data was uploaded to the PRIDE Archive and is publicly available through the following identifiers: PXD011235 (COMA-Sli15/Ipl1); PXD011236 (CCAN)

The following data sets were generated
    1. Solis V et al
    (2019) AOCM-CPC
    Pride Archive, PXD011235 (COMA-Sli15/Ipl1).

Article and author information

Author details

  1. Josef Fischböck-Halwachs

    Gene Center Munich, Ludwig-Maximilians-Universität München, Munich, Germany
    Competing interests
    The authors declare that no competing interests exist.
  2. Sylvia Singh

    Gene Center Munich, Ludwig-Maximilians-Universität München, Munich, Germany
    Competing interests
    The authors declare that no competing interests exist.
  3. Mia Potocnjak

    Gene Center Munich, Ludwig-Maximilians-Universität München, Munich, Germany
    Competing interests
    The authors declare that no competing interests exist.
  4. Götz Hagemann

    Gene Center Munich, Ludwig-Maximilians-Universität München, Munich, Germany
    Competing interests
    The authors declare that no competing interests exist.
  5. Victor Solis-Mezarino

    Gene Center Munich, Ludwig-Maximilians-Universität München, Munich, Germany
    Competing interests
    The authors declare that no competing interests exist.
  6. Stephan Woike

    Gene Center, Ludwig-Maximilians-Universität München, Munich, Germany
    Competing interests
    The authors declare that no competing interests exist.
  7. Medini Ghodgaonkar Steger

    Gene Center Munich, Ludwig-Maximilians-Universität München, Munich, Germany
    Competing interests
    The authors declare that no competing interests exist.
  8. Florian Weissmann

    Research Institute of Molecular Pathology, Vienna Biocenter, Vienna, Germany
    Competing interests
    The authors declare that no competing interests exist.
  9. Laura D Gallego

    Max F Perutz Laboratories, Medical University of Vienna, Vienna, Austria
    Competing interests
    The authors declare that no competing interests exist.
  10. Julie Rojas

    Laboratory of Chromosome Biology, Max Planck Institute of Biochemistry, Martinsried, Germany
    Competing interests
    The authors declare that no competing interests exist.
  11. Jessica Andreani-Feuillet

    Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, University Paris-Sud, Gif-sur-Yvette, France
    Competing interests
    The authors declare that no competing interests exist.
  12. Alwin Köhler

    Max F Perutz Laboratories, Medical University of Vienna, Vienna, Austria
    Competing interests
    The authors declare that no competing interests exist.
  13. Franz Herzog

    Gene Center Munich, Ludwig-Maximilians-Universität München, Munich, Germany
    For correspondence
    herzog@genzentrum.lmu.de
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8270-1449

Funding

European Research Council (ERC-StG MolStruKT no 638218)

  • Franz Herzog

Human Frontier Science Program (RGP0008/2015)

  • Franz Herzog

Deutsche Forschungsgemeinschaft

  • Mia Potocnjak
  • Götz Hagemann
  • Victor Solis-Mezarino
  • Franz Herzog

Bavarian Research Center for Molecular Biosystems

  • Franz Herzog

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

Reviewing Editor

  1. Jennifer G. DeLuca, Colorado State University, United States

Publication history

  1. Received: October 16, 2018
  2. Accepted: May 20, 2019
  3. Accepted Manuscript published: May 21, 2019 (version 1)
  4. Version of Record published: June 3, 2019 (version 2)

Copyright

© 2019, Fischböck-Halwachs 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.

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  1. Josef Fischböck-Halwachs
  2. Sylvia Singh
  3. Mia Potocnjak
  4. Götz Hagemann
  5. Victor Solis-Mezarino
  6. Stephan Woike
  7. Medini Ghodgaonkar Steger
  8. Florian Weissmann
  9. Laura D Gallego
  10. Julie Rojas
  11. Jessica Andreani-Feuillet
  12. Alwin Köhler
  13. Franz Herzog
(2019)
The COMA complex interacts with Cse4 and positions Sli15/Ipl1 at the budding yeast inner kinetochore
eLife 8:e42879.
https://doi.org/10.7554/eLife.42879

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