Selective dephosphorylation by PP2A-B55 directs the meiosis I - meiosis II transition in oocytes

  1. S Zachary Swartz
  2. Hieu T Nguyen
  3. Brennan C McEwan
  4. Mark E Adamo
  5. Iain M Cheeseman  Is a corresponding author
  6. Arminja N Kettenbach  Is a corresponding author
  1. Massachusetts Institute of Technology, United States
  2. Geisel School of Medicine at Dartmouth, United States
  3. Whitehead Institute, United States

Abstract

Meiosis is a specialized cell cycle that requires sequential changes to the cell division machinery to facilitate changing functions. To define the mechanisms that enable the oocyte-to-embryo transition, we performed time-course proteomics in synchronized sea star oocytes from prophase I through the first embryonic cleavage. Although we find that protein levels are broadly stable, our analysis reveals that dynamic waves of phosphorylation underlie each meiotic stage. We find that the phosphatase PP2A-B55 is reactivated at the meiosis I/II transition resulting in the preferential dephosphorylation of threonine residues. Selective dephosphorylation is critical for directing the MI / MII transition as altering PP2A-B55 substrate preferences disrupts key cell cycle events after meiosis I. In addition, threonine to serine substitution of a conserved phosphorylation site in the substrate INCENP prevents its relocalization at anaphase I. Thus, through its inherent phospho-threonine preference, PP2A-B55 imposes specific phosphoregulated behaviors that distinguish the two meiotic divisions.

Data availability

Raw MS data for the experiments performed in this study are available at MassIVE and ProteomeXchange, accession number: PXD020916, password: p845. Plasmids generated from this study are deposited to Addgene. Custom R script is available at Github.

The following data sets were generated

Article and author information

Author details

  1. S Zachary Swartz

    Biology, Massachusetts Institute of Technology, Cambridge, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Hieu T Nguyen

    Department of Biochemistry and Cell Biology, Geisel School of Medicine at Dartmouth, Hanover, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Brennan C McEwan

    Department of Biochemistry and Cell Biology, Geisel School of Medicine at Dartmouth, Hanover, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Mark E Adamo

    Norris Cotton Cancer Center, Geisel School of Medicine at Dartmouth, Lebanon, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Iain M Cheeseman

    Department of Biology, MIT, Whitehead Institute, Cambridge, United States
    For correspondence
    icheese@wi.mit.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3829-5612
  6. Arminja N Kettenbach

    Department of Biochemistry and Cell Biology, Geisel School of Medicine at Dartmouth, Lebanon, United States
    For correspondence
    Arminja.N.Kettenbach@dartmouth.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3979-4576

Funding

National Institute of General Medical Sciences (R35GM126930)

  • Iain M Cheeseman

National Institute of General Medical Sciences (R35GM119455)

  • Arminja N Kettenbach

Eunice Kennedy Shriver National Institute of Child Health and Human Development (K99HD099315)

  • S Zachary Swartz

Gordon and Betty Moore Foundation

  • Iain M Cheeseman

Global Consortium for Reproductive Longevity and Equity (GCRLE-1220)

  • Iain M Cheeseman

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

Copyright

© 2021, Swartz 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.

Metrics

  • 1,801
    views
  • 281
    downloads
  • 14
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. S Zachary Swartz
  2. Hieu T Nguyen
  3. Brennan C McEwan
  4. Mark E Adamo
  5. Iain M Cheeseman
  6. Arminja N Kettenbach
(2021)
Selective dephosphorylation by PP2A-B55 directs the meiosis I - meiosis II transition in oocytes
eLife 10:e70588.
https://doi.org/10.7554/eLife.70588

Share this article

https://doi.org/10.7554/eLife.70588

Further reading

    1. Cell Biology
    Jessica E Schwarz, Antonijo Mrčela ... Amita Sehgal
    Short Report

    Aging is associated with a number of physiologic changes including perturbed circadian rhythms; however, mechanisms by which rhythms are altered remain unknown. To test the idea that circulating factors mediate age-dependent changes in peripheral rhythms, we compared the ability of human serum from young and old individuals to synchronize circadian rhythms in culture. We collected blood from apparently healthy young (age 25–30) and old (age 70–76) individuals at 14:00 and used the serum to synchronize cultured fibroblasts. We found that young and old sera are equally competent at initiating robust ~24 hr oscillations of a luciferase reporter driven by clock gene promoter. However, cyclic gene expression is affected, such that young and old sera promote cycling of different sets of genes. Genes that lose rhythmicity with old serum entrainment are associated with oxidative phosphorylation and Alzheimer’s Disease as identified by STRING and IPA analyses. Conversely, the expression of cycling genes associated with cholesterol biosynthesis increased in the cells entrained with old serum. Genes involved in the cell cycle and transcription/translation remain rhythmic in both conditions. We did not observe a global difference in the distribution of phase between groups, but found that peak expression of several clock-controlled genes (PER3, NR1D1, NR1D2, CRY1, CRY2, and TEF) lagged in the cells synchronized ex vivo with old serum. Taken together, these findings demonstrate that age-dependent blood-borne factors affect circadian rhythms in peripheral cells and have the potential to impact health and disease via maintaining or disrupting rhythms respectively.

    1. Cell Biology
    Ewa K Bomba-Warczak, Karen M Velez ... Francesca E Duncan
    Short Report

    The mechanisms contributing to age-related deterioration of the female reproductive system are complex, however aberrant protein homeostasis is a major contributor. We elucidated exceptionally stable proteins, structures, and macromolecules that persist in mammalian ovaries and gametes across the reproductive lifespan. Ovaries exhibit localized structural and cell-type-specific enrichment of stable macromolecules in both the follicular and extrafollicular environments. Moreover, ovaries and oocytes both harbor a panel of exceptionally long-lived proteins, including cytoskeletal, mitochondrial, and oocyte-derived proteins. The exceptional persistence of these long-lived molecules suggest a critical role in lifelong maintenance and age-dependent deterioration of reproductive tissues.