Differential regulation of the proteome and phosphosproteome along the dorso-ventral axis of the early Drosophila embryo

  1. Juan Manuel Gomez
  2. Hendrik Nolte
  3. Elisabeth Vogelsang
  4. Bipasha Dey
  5. Michiko Takeda
  6. Girolamo Giudice
  7. Miriam Faxel
  8. Theresa Haunold
  9. Alina Cepraga
  10. Robert P Zinzen
  11. Marcus Krüger
  12. Evangelia Petsalaki
  13. Yu-Chiun Wang
  14. Maria Leptin  Is a corresponding author
  1. European Molecular Biology Laboratory, Germany
  2. University of Cologne, Germany
  3. RIKEN Center for Biosystems Dynamics Research, Japan
  4. European Molecular Biology Laboratory, United Kingdom
  5. Max Delbrück Center for Molecular Medicine, Germany

Abstract

The initially homogeneous epithelium of the early Drosophila embryo differentiates into regional subpopulations with different behaviours and physical properties that are needed for morphogenesis. The factors at top of the genetic hierarchy that control these behaviours are known, but many of their targets are not. To understand how proteins work together to mediate differential cellular activities, we studied in an unbiased manner the proteomes and phosphoproteomes of the three main cell populations along the dorso-ventral axis during gastrulation using mutant embryos that represent the different populations. We detected 6111 protein groups and 6259 phosphosites of which 3398 and 3433 respectively, were differentially regulated. The changes in phosphosite abundance did not correlate with changes in host protein abundance, showing phosphorylation to be a regulatory step during gastrulation. Hierarchical clustering of protein groups and phosphosites identified clusters that contain known fate determinants such as Doc1, Sog, Snail and Twist. The recovery of the appropriate known marker proteins in each of the different mutants we used validated the approach, but also revealed that two mutations that both interfere with the dorsal fate pathway, Toll10B and serpin27aex do this in very different manners. Diffused network analyses within each cluster point to microtubule components as one of the main groups of regulated proteins. Functional studies on the role of microtubules provide the proof of principle that microtubules have different functions in different domains along the DV axis of the embryo.

Data availability

The whole proteome and phosphoproteomic data is available.The raw files for the proteomics and phosphoproteomics experiments were deposited in PRIDE under separate identifiers:Proteome: Identifier PXD046050Phosphoproteome: Identifier PXD046192

The following data sets were generated
The following previously published data sets were used

Article and author information

Author details

  1. Juan Manuel Gomez

    Directors's Research and Developmental Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  2. Hendrik Nolte

    CECAD Research Center, University of Cologne, Cologne, Germany
    Competing interests
    The authors declare that no competing interests exist.
  3. Elisabeth Vogelsang

    Institute of Genetics, University of Cologne, Cologne, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6817-5953
  4. Bipasha Dey

    RIKEN Center for Biosystems Dynamics Research, Kobe, Japan
    Competing interests
    The authors declare that no competing interests exist.
  5. Michiko Takeda

    RIKEN Center for Biosystems Dynamics Research, Kobe, Japan
    Competing interests
    The authors declare that no competing interests exist.
  6. Girolamo Giudice

    European Bioinformatics Institute, European Molecular Biology Laboratory, Hinxton, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5359-8208
  7. Miriam Faxel

    Max Delbrück Center for Molecular Medicine, Berlin, Germany
    Competing interests
    The authors declare that no competing interests exist.
  8. Theresa Haunold

    Director's Research and Developmental Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0009-0007-8343-4945
  9. Alina Cepraga

    Director's Research and Developmental Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0009-0004-6161-1195
  10. Robert P Zinzen

    Max Delbrück Center for Molecular Medicine, Berlin, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8638-5102
  11. Marcus Krüger

    CECAD Research Center, University of Cologne, Cologne, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5846-6941
  12. Evangelia Petsalaki

    European Bioinformatics Institute, European Molecular Biology Laboratory, Hinxton, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8294-2995
  13. Yu-Chiun Wang

    RIKEN Center for Biosystems Dynamics Research, Kobe, Japan
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3797-4138
  14. Maria Leptin

    Institute for Genetics, University of Cologne, Cologne, Germany
    For correspondence
    mleptin@uni-koeln.de
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7097-348X

Funding

European Molecular Biology Organization (N/A)

  • Maria Leptin

Deutsche Forschungsgemeinschaft (LE 546/12)

  • Maria Leptin

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

Copyright

© 2024, Gomez 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. Juan Manuel Gomez
  2. Hendrik Nolte
  3. Elisabeth Vogelsang
  4. Bipasha Dey
  5. Michiko Takeda
  6. Girolamo Giudice
  7. Miriam Faxel
  8. Theresa Haunold
  9. Alina Cepraga
  10. Robert P Zinzen
  11. Marcus Krüger
  12. Evangelia Petsalaki
  13. Yu-Chiun Wang
  14. Maria Leptin
(2024)
Differential regulation of the proteome and phosphosproteome along the dorso-ventral axis of the early Drosophila embryo
eLife 13:e99263.
https://doi.org/10.7554/eLife.99263

Share this article

https://doi.org/10.7554/eLife.99263

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