Differential regulation of the proteome and phosphosproteome along the dorso-ventral axis of the early Drosophila embryo
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
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Single Cell RNAseq Atlas - Drosophila gastrulationNCBI Gene Expression Omnibus, GSE95025.
Article and author information
Author details
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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