Abstract

The plant corepressor TOPLESS (TPL) is recruited to a large number of loci that are selectively induced in response to developmental or environmental cues, yet the mechanisms by which it inhibits expression in the absence of these stimuli is poorly understood. Previously, we had used the N-terminus of Arabidopsis thaliana TPL to enable repression of a synthetic auxin response circuit in Saccharomyces cerevisiae (yeast). Here, we leveraged the yeast system to interrogate the relationship between TPL structure and function, specifically scanning for repression domains. We identified a potent repression domain in Helix 8 located within the CRA domain, which directly interacted with the Mediator middle module subunits Med21 and Med10. Interactions between TPL and Mediator were required to fully repress transcription in both yeast and plants. In contrast, we found that multimer formation, a conserved feature of many corepressors, had minimal influence on the repression strength of TPL.

Data availability

All flow cytometry data is deposited at https://flowrepository.org/. Repository IDS: FR-FCM-Z2GM, FR-FCM-Z2GR, FR-FCM-Z2GQ, FR-FCM-Z2GX, FR-FCM-Z2GT, FR-FCM-Z2W2. All protein interactions have been deposited to IntAct, under the accession code, IM-28972. All code is available through Github: https://github.com/achillobator/TPL_Structure_Function/

The following data sets were generated

Article and author information

Author details

  1. Alexander R Leydon

    Biology, University of Washington, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3034-1482
  2. Wei Wang

    Pharmacology, University of Washington, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Hardik P Gala

    Biology, University of Washington, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Sabrina Gilmour

    Biology, University of Washington, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Samuel Juarez-Solis

    Biology, University of Washington, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Mollye L Zahler

    Biology, University of Washington, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Joseph E Zemke

    Biology, University of Washington, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. NIng Zheng

    Pharmacology, University of Washington, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Jennifer L Nemhauser

    Department of Biology, University of Washington, Seattle, United States
    For correspondence
    jn7@uw.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8909-735X

Funding

National Institutes of Health (GM107084)

  • Alexander R Leydon
  • Hardik P Gala
  • Sabrina Gilmour
  • Samuel Juarez-Solis
  • Mollye L Zahler
  • Joseph E Zemke
  • Jennifer L Nemhauser

Howard Hughes Medical Institute

  • Alexander R Leydon
  • Wei Wang
  • Hardik P Gala
  • Sabrina Gilmour
  • Samuel Juarez-Solis
  • Mollye L Zahler
  • Joseph E Zemke
  • NIng Zheng
  • Jennifer L Nemhauser

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

Reviewing Editor

  1. Irwin Davidson, Institut de Génétique et de Biologie Moléculaire et Cellulaire, France

Version history

  1. Received: January 21, 2021
  2. Accepted: May 31, 2021
  3. Accepted Manuscript published: June 2, 2021 (version 1)
  4. Version of Record published: June 14, 2021 (version 2)

Copyright

© 2021, Leydon 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. Alexander R Leydon
  2. Wei Wang
  3. Hardik P Gala
  4. Sabrina Gilmour
  5. Samuel Juarez-Solis
  6. Mollye L Zahler
  7. Joseph E Zemke
  8. NIng Zheng
  9. Jennifer L Nemhauser
(2021)
Repression by the Arabidopsis TOPLESS corepressor requires association with the core Mediator complex
eLife 10:e66739.
https://doi.org/10.7554/eLife.66739

Share this article

https://doi.org/10.7554/eLife.66739

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