Mitochondrial copper and phosphate transporter specificity was defined early in the evolution of eukaryotes

  1. Xinyu Zhu
  2. Aren Boulet
  3. Katherine M Buckley
  4. Casey B Phillips
  5. Micah G Gammon
  6. Laura E Oldfather
  7. Stanley A Moore
  8. Scot C Leary
  9. Paul A Cobine  Is a corresponding author
  1. Auburn University, United States
  2. University of Saskatchewan, Canada
  3. University Saskatchewan, Canada

Abstract

The mitochondrial carrier family protein SLC25A3 transports both copper and phosphate in mammals yet in Saccharomyces cerevisiae the transport of these substrates is partitioned across two paralogs: PIC2 and MIR1. To understand the ancestral state of copper and phosphate transport in mitochondria, we explored the evolutionary relationships of PIC2 and MIR1 orthologs across the eukaryotic tree of life. Phylogenetic analyses revealed that PIC2-like and MIR1-like orthologs are present in all major eukaryotic supergroups, indicating an ancient gene duplication created these paralogs. To link this phylogenetic signal to protein function, we used structural modelling and site-directed mutagenesis to identify residues involved in copper and phosphate transport. Based on these analyses, we generated a L175A variant of mouse SLC25A3 that retains the ability to transport copper but not phosphate. This work highlights the utility of using an evolutionary framework to uncover amino acids involved in substrate recognition by mitochondrial carrier family proteins.

Data availability

All data generated or analyzed during this study are included in the manuscript, supplemental file, and available on GenBank.

The following previously published data sets were used
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Article and author information

Author details

  1. Xinyu Zhu

    Biological Sciences, Auburn University, Auburn, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Aren Boulet

    Biochemistry, Microbiology and Immunology, University of Saskatchewan, Saskatoon, Canada
    Competing interests
    The authors declare that no competing interests exist.
  3. Katherine M Buckley

    Biological Sciences, Auburn University, Auburn, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Casey B Phillips

    Biological Sciences, Auburn University, Auburn, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Micah G Gammon

    Biological Sciences, Auburn University, Auburn, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Laura E Oldfather

    Biological Sciences, Auburn University, Auburn, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Stanley A Moore

    Biochemistry, Microbiology and Immunology, University Saskatchewan, Saskatoon, Canada
    Competing interests
    The authors declare that no competing interests exist.
  8. Scot C Leary

    Biochemistry, Microbiology and Immunology, University of Saskatchewan, Saskatoon, Canada
    Competing interests
    The authors declare that no competing interests exist.
  9. Paul A Cobine

    Biological Sciences, Auburn University, Auburn, United States
    For correspondence
    paul.cobine@auburn.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6012-0985

Funding

National Institutes of Health (R01GM120211)

  • Scot C Leary
  • Paul A Cobine

National Science Foundation (EF 2021886)

  • Katherine M Buckley

Alabama Agricultural Experiment Station

  • Paul A Cobine

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

Reviewing Editor

  1. Randy B Stockbridge, University of Michigan, United States

Version history

  1. Received: November 7, 2020
  2. Accepted: February 15, 2021
  3. Accepted Manuscript published: February 16, 2021 (version 1)
  4. Version of Record published: March 2, 2021 (version 2)

Copyright

© 2021, Zhu 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. Xinyu Zhu
  2. Aren Boulet
  3. Katherine M Buckley
  4. Casey B Phillips
  5. Micah G Gammon
  6. Laura E Oldfather
  7. Stanley A Moore
  8. Scot C Leary
  9. Paul A Cobine
(2021)
Mitochondrial copper and phosphate transporter specificity was defined early in the evolution of eukaryotes
eLife 10:e64690.
https://doi.org/10.7554/eLife.64690

Share this article

https://doi.org/10.7554/eLife.64690

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