SILAC-based quantitative proteomic analysis of Drosophila gastrula stage embryos mutant for fibroblast growth factor signalling.
Fly (Austin)
; 14(1-4): 10-28, 2020.
Article
em En
| MEDLINE
| ID: mdl-31873056
ABSTRACT
Quantitative proteomic analyses in combination with genetics provide powerful tools in developmental cell signalling research. Drosophila melanogaster is one of the most widely used genetic models for studying development and disease. Here we combined quantitative proteomics with genetic selection to determine changes in the proteome upon depletion of Heartless (Htl) Fibroblast-Growth Factor (FGF) receptor signalling in Drosophila embryos at the gastrula stage. We present a robust, single generation SILAC (stable isotope labelling with amino acids in cell culture) protocol for labelling proteins in early embryos. For the selection of homozygously mutant embryos at the pre-gastrula stage, we developed an independent genetic marker. Our analyses detected quantitative changes in the global proteome of htl mutant embryos during gastrulation. We identified distinct classes of downregulated and upregulated proteins, and network analyses indicate functionally related groups of proteins in each class. In addition, we identified changes in the abundance of phosphopeptides. In summary, our quantitative proteomic analysis reveals global changes in metabolic, nucleoplasmic, cytoskeletal and transport proteins in htl mutant embryos.
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MEDLINE
Assunto principal:
Proteínas Tirosina Quinases
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Receptores de Fatores de Crescimento de Fibroblastos
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Regulação da Expressão Gênica no Desenvolvimento
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Proteínas de Drosophila
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Drosophila melanogaster
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Fatores de Crescimento de Fibroblastos
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Gástrula
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article