Quality control in oocytes by p63 is based on a spring-loaded activation mechanism on the molecular and cellular level

  1. Daniel Coutandin
  2. Christian Osterburg
  3. Ratnesh Kumar Srivastav
  4. Manuela Sumyk
  5. Sebastian Kehrloesser
  6. Jakob Gebel
  7. Marcel Tuppi
  8. Jens Hannewald
  9. Birgit Schäfer
  10. Eidarus Salah
  11. Sebastian Mathea
  12. Uta Müller-Kuller
  13. James Doutch
  14. Manuel Grez
  15. Stefan Knapp
  16. Volker Dötsch  Is a corresponding author
  1. Goethe University, Germany
  2. Merck KGaA, Germany
  3. University of Oxford, United Kingdom
  4. Georg-Speyer Haus, Germany
  5. ISIS Neutron and Muon Source, United Kingdom
  6. Georg-Speyer-Haus, Germany

Abstract

Mammalian oocytes are arrested in the dictyate stage of meiotic prophase I for long periods of time, during which the high concentration of the p53 family member TAp63α sensitizes them to DNA damage-induced apoptosis. TAp63α is kept in an inactive and exclusively dimeric state but undergoes rapid phosphorylation-induced tetramerization and concomitant activation upon detection of DNA damage. Here we show that the TAp63α dimer is a kinetically trapped state. Activation follows a spring-loaded mechanism not requiring further translation of other cellular factors in oocytes and is associated with unfolding of the inhibitory structure that blocks the tetramerization interface. Using a combination of biophysical methods as well as cell and ovary culture experiments we explain how TAp63α is kept inactive in the absence of DNA damage but causes rapid oocyte elimination in response to a few DNA double strand breaks thereby acting as the key quality control factor in maternal reproduction.

Article and author information

Author details

  1. Daniel Coutandin

    Institute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance and Cluster of Excellence Macromolecular Complexes, Goethe University, Frankfurt, Germany
    Competing interests
    No competing interests declared.
  2. Christian Osterburg

    Institute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance and Cluster of Excellence Macromolecular Complexes, Goethe University, Frankfurt, Germany
    Competing interests
    No competing interests declared.
  3. Ratnesh Kumar Srivastav

    Institute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance and Cluster of Excellence Macromolecular Complexes, Goethe University, Frankfurt, Germany
    Competing interests
    No competing interests declared.
  4. Manuela Sumyk

    Institute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance and Cluster of Excellence Macromolecular Complexes, Goethe University, Frankfurt, Germany
    Competing interests
    No competing interests declared.
  5. Sebastian Kehrloesser

    Institute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance and Cluster of Excellence Macromolecular Complexes, Goethe University, Frankfurt, Germany
    Competing interests
    No competing interests declared.
  6. Jakob Gebel

    Institute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance and Cluster of Excellence Macromolecular Complexes, Goethe University, Frankfurt, Germany
    Competing interests
    No competing interests declared.
  7. Marcel Tuppi

    Institute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance and Cluster of Excellence Macromolecular Complexes, Goethe University, Frankfurt, Germany
    Competing interests
    No competing interests declared.
  8. Jens Hannewald

    MS-DTB-C Protein Purification, Merck KGaA, Darmstadt, Germany
    Competing interests
    No competing interests declared.
  9. Birgit Schäfer

    Institute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance and Cluster of Excellence Macromolecular Complexes, Goethe University, Frankfurt, Germany
    Competing interests
    No competing interests declared.
  10. Eidarus Salah

    Nuffield Department of Medicine, Structural Genomics Consortium, University of Oxford, Oxford, United Kingdom
    Competing interests
    No competing interests declared.
  11. Sebastian Mathea

    Nuffield Department of Medicine, Structural Genomics Consortium, University of Oxford, Oxford, United Kingdom
    Competing interests
    No competing interests declared.
  12. Uta Müller-Kuller

    Georg-Speyer Haus, Frankfurt, Germany
    Competing interests
    No competing interests declared.
  13. James Doutch

    Rutherford Appleton Laboratory, ISIS Neutron and Muon Source, Dodcot, United Kingdom
    Competing interests
    No competing interests declared.
  14. Manuel Grez

    Georg-Speyer-Haus, Frankfurt, Germany
    Competing interests
    No competing interests declared.
  15. Stefan Knapp

    Nuffield Department of Medicine, Structural Genomics Consortium, University of Oxford, Oxford, United Kingdom
    Competing interests
    No competing interests declared.
  16. Volker Dötsch

    Institute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance and Cluster of Excellence Macromolecular Complexes, Goethe University, Frankfurt, Germany
    For correspondence
    vdoetsch@em.uni-frankfurt.de
    Competing interests
    Volker Dötsch, Reviewing editor, eLife.

Reviewing Editor

  1. Joaquín M Espinosa, University of Colorado Denver School of Medicine, United States

Ethics

Animal experimentation: The work with mice was conducted according to the regulations of the Goethe University and the DFG (according to {section sign} 4 TierSchG) and supervised by the Tierschutzbeauftragte of Goethe University.

Version history

  1. Received: December 18, 2015
  2. Accepted: March 28, 2016
  3. Accepted Manuscript published: March 29, 2016 (version 1)
  4. Version of Record published: April 29, 2016 (version 2)

Copyright

© 2016, Coutandin 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

  • 2,326
    views
  • 470
    downloads
  • 51
    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. Daniel Coutandin
  2. Christian Osterburg
  3. Ratnesh Kumar Srivastav
  4. Manuela Sumyk
  5. Sebastian Kehrloesser
  6. Jakob Gebel
  7. Marcel Tuppi
  8. Jens Hannewald
  9. Birgit Schäfer
  10. Eidarus Salah
  11. Sebastian Mathea
  12. Uta Müller-Kuller
  13. James Doutch
  14. Manuel Grez
  15. Stefan Knapp
  16. Volker Dötsch
(2016)
Quality control in oocytes by p63 is based on a spring-loaded activation mechanism on the molecular and cellular level
eLife 5:e13909.
https://doi.org/10.7554/eLife.13909

Share this article

https://doi.org/10.7554/eLife.13909

Further reading

    1. Structural Biology and Molecular Biophysics
    Hitendra Negi, Aravind Ravichandran ... Ranabir Das
    Research Article

    The proteasome controls levels of most cellular proteins, and its activity is regulated under stress, quiescence, and inflammation. However, factors determining the proteasomal degradation rate remain poorly understood. Proteasome substrates are conjugated with small proteins (tags) like ubiquitin and Fat10 to target them to the proteasome. It is unclear if the structural plasticity of proteasome-targeting tags can influence substrate degradation. Fat10 is upregulated during inflammation, and its substrates undergo rapid proteasomal degradation. We report that the degradation rate of Fat10 substrates critically depends on the structural plasticity of Fat10. While the ubiquitin tag is recycled at the proteasome, Fat10 is degraded with the substrate. Our results suggest significantly lower thermodynamic stability and faster mechanical unfolding in Fat10 compared to ubiquitin. Long-range salt bridges are absent in the Fat10 structure, creating a plastic protein with partially unstructured regions suitable for proteasome engagement. Fat10 plasticity destabilizes substrates significantly and creates partially unstructured regions in the substrate to enhance degradation. NMR-relaxation-derived order parameters and temperature dependence of chemical shifts identify the Fat10-induced partially unstructured regions in the substrate, which correlated excellently to Fat10-substrate contacts, suggesting that the tag-substrate collision destabilizes the substrate. These results highlight a strong dependence of proteasomal degradation on the structural plasticity and thermodynamic properties of the proteasome-targeting tags.

    1. Biochemistry and Chemical Biology
    2. Structural Biology and Molecular Biophysics
    Amy H Andreotti, Volker Dötsch
    Editorial

    The articles in this special issue highlight how modern cellular, biochemical, biophysical and computational techniques are allowing deeper and more detailed studies of allosteric kinase regulation.