Abstract

Cellular redox status affects diverse cellular functions, including proliferation, protein homeostasis, and aging. Thus, individual differences in redox status can give rise to distinct sub-populations even among cells with identical genetic backgrounds. Here, we have created a novel methodology to track redox status at single cell resolution using the redox-sensitive probe Grx1-roGFP2. Our method allows identification and sorting of sub-populations with different oxidation levels in either the cytosol, mitochondria or peroxisomes. Using this approach, we defined a redox-dependent heterogeneity of yeast cells and characterized growth, as well as proteomic and transcriptomic profiles of distinctive redox subpopulations. We report that, starting in late logarithmic growth, cells of the same age have a bi-modal distribution of oxidation status. A comparative proteomic analysis between these populations identified three key proteins, Hsp30, Dhh1, and Pnc1, which affect basal oxidation levels and may serve as first line of defense proteins in redox homeostasis.

Data availability

All data generated or analyses during this study are included in the manuscript and supporting files. Proteomic data was uploadedto the PRIDE database with the dataset identifier PXD009443. Transcriptomic data was uploaded to the GEO database as described in the manuscript (methods).

The following data sets were generated

Article and author information

Author details

  1. Meytal Radzinski

    Department of Biological Chemistry, The Hebrew University of Jerusalem, Jerusalem, Israel
    Competing interests
    The authors declare that no competing interests exist.
  2. Rosi Fassler

    Department of Biological Chemistry, The Hebrew University of Jerusalem, Jerusalem, Israel
    Competing interests
    The authors declare that no competing interests exist.
  3. Ohad Yogev

    Department of Biological Chemistry, The Hebrew University of Jerusalem, Jerusalem, Israel
    Competing interests
    The authors declare that no competing interests exist.
  4. William Breuer

    Proteomics and Mass Spectrometry Unit, The Hebrew University of Jerusalem, Jerusalem, Israel
    Competing interests
    The authors declare that no competing interests exist.
  5. Nadav Shai

    Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2812-3884
  6. Jenia Gutin

    Department of Biological Chemistry, The Hebrew University of Jerusalem, Jerusalem, Israel
    Competing interests
    The authors declare that no competing interests exist.
  7. Sidra Ilyas

    Department of Biological Chemistry, The Hebrew University of Jerusalem, Jerusalem, Israel
    Competing interests
    The authors declare that no competing interests exist.
  8. Yifat Geffen

    Department of Biological Chemistry, The Hebrew University of Jerusalem, Jerusalem, Israel
    Competing interests
    The authors declare that no competing interests exist.
  9. Sabina Tsytkin-Kirschenzweig

    Grass Center for Bioengineering, Benin School of Computer Science and Engineering, The Hebrew University of Jerusalem, Jerusalem, Israel
    Competing interests
    The authors declare that no competing interests exist.
  10. Yaakov Nahmias

    Grass Center for Bioengineering, Benin School of Computer Science and Engineering, The Hebrew University of Jerusalem, Jerusalem, Israel
    Competing interests
    The authors declare that no competing interests exist.
  11. Tommer Ravid

    Department of Biological Chemistry, The Hebrew University of Jerusalem, Jerusalem, Israel
    Competing interests
    The authors declare that no competing interests exist.
  12. Nir Friedman

    Department of Biological Chemistry, The Hebrew University of Jerusalem, Jerusalem, Israel
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9678-3550
  13. Maya Shuldiner

    Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel
    Competing interests
    The authors declare that no competing interests exist.
  14. Dana Reichmann

    Department of Biological Chemistry, The Hebrew University of Jerusalem, Jerusalem, Israel
    For correspondence
    danare@mail.huji.ac.il
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0315-5334

Funding

Israel Science Foundation

  • Dana Reichmann

Human Frontier Science Program

  • Dana Reichmann

European Commission

  • Dana Reichmann

US-Binational Science Foundation

  • Dana Reichmann

Joint Berlin-Jerusalem postdoc fellowship, Hebrew University and Freie University

  • Sidra Ilyas

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

Copyright

© 2018, Radzinski 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. Meytal Radzinski
  2. Rosi Fassler
  3. Ohad Yogev
  4. William Breuer
  5. Nadav Shai
  6. Jenia Gutin
  7. Sidra Ilyas
  8. Yifat Geffen
  9. Sabina Tsytkin-Kirschenzweig
  10. Yaakov Nahmias
  11. Tommer Ravid
  12. Nir Friedman
  13. Maya Shuldiner
  14. Dana Reichmann
(2018)
Temporal profiling of redox-dependent heterogeneity in single cells
eLife 7:e37623.
https://doi.org/10.7554/eLife.37623

Share this article

https://doi.org/10.7554/eLife.37623

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