1. Genetics and Genomics
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Genetic profiling of protein burden and nuclear export overload

  1. Reiko Kintaka
  2. Koji Makanae
  3. Shotaro Namba
  4. Hisaaki Kato
  5. Keiji Kito
  6. Shinsuke Ohnuki
  7. Yoshikazu Ohya
  8. Brenda J Andrews
  9. Charles Boone
  10. Hisao Moriya  Is a corresponding author
  1. University of Toronto, Canada
  2. Okayama University, Japan
  3. Meiji University, Japan
  4. University of Tokyo, Japan
Research Article
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Cite this article as: eLife 2020;9:e54080 doi: 10.7554/eLife.54080

Abstract

Overproduction (op) of proteins triggers cellular defects. One of the consequences of overproduction is the protein burden/cost, which is produced by an overloading of the protein synthesis process. However, the physiology of cells under a protein burden is not well characterized. We performed genetic profiling of protein burden by systematic analysis of genetic interactions between GFP-op, surveying both deletion and temperature-sensitive mutants in budding yeast. We also performed genetic profiling in cells with overproduction of triple-GFP (tGFP), and the nuclear export signal-containing tGFP (NES-tGFP). The mutants specifically interacted with GFP-op were suggestive of unexpected connections between actin-related processes like polarization and the protein burden, which was supported by morphological analysis. The tGFP-op interactions suggested that this protein probe overloads the proteasome, whereas those that interacted with NES-tGFP involved genes encoding components of the nuclear export process, providing a resource for further analysis of the protein burden and nuclear export overload.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files.

Article and author information

Author details

  1. Reiko Kintaka

    Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Canada
    Competing interests
    The authors declare that no competing interests exist.
  2. Koji Makanae

    Research Core for Interdisciplinary Sciences, Okayama University, Kita-ku, Okayama, Japan
    Competing interests
    The authors declare that no competing interests exist.
  3. Shotaro Namba

    Matching Program Course, Okayama University, Okayama, Japan
    Competing interests
    The authors declare that no competing interests exist.
  4. Hisaaki Kato

    Graduate School of Environmental and Life Science, Okayama University, Okayama, Japan
    Competing interests
    The authors declare that no competing interests exist.
  5. Keiji Kito

    Department of Life Sciences, School of Agriculture, Meiji University, Kawasaki, Japan
    Competing interests
    The authors declare that no competing interests exist.
  6. Shinsuke Ohnuki

    Graduate School of Frontier Sciences, University of Tokyo, Kashiwa, Japan
    Competing interests
    The authors declare that no competing interests exist.
  7. Yoshikazu Ohya

    Graduate School of Frontier Sciences, University of Tokyo, Kashiwa, Japan
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0837-1239
  8. Brenda J Andrews

    Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Canada
    Competing interests
    The authors declare that no competing interests exist.
  9. Charles Boone

    Donnelly Center for Cellular and Biomolecular Research, University of Toronto, Toronto, Canada
    Competing interests
    The authors declare that no competing interests exist.
  10. Hisao Moriya

    Research Core for Interdisciplinary Sciences, Okayama University, Kita-ku, Okayama, Japan
    For correspondence
    hisaom@cc.okayama-u.ac.jp
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7638-3640

Funding

Japan Society for the Promotion of Science (17H03618)

  • Hisao Moriya

Japan Society for the Promotion of Science (15KK0258)

  • Hisao Moriya

Japan Society for the Promotion of Science (18K19300)

  • Hisao Moriya

Japan Society for the Promotion of Science (20H03242)

  • Hisao Moriya

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

Reviewing Editor

  1. Nir Ben-Tal, Tel Aviv University, Israel

Publication history

  1. Received: December 2, 2019
  2. Accepted: November 1, 2020
  3. Accepted Manuscript published: November 4, 2020 (version 1)
  4. Version of Record published: November 18, 2020 (version 2)

Copyright

© 2020, Kintaka 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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