SILAC-based quantitative proteomic analysis of Drosophila gastrula stage embryos mutant for fibroblast growth factor signalling
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 t...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2020-10-01
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| Series: | Fly |
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| Online Access: | http://dx.doi.org/10.1080/19336934.2019.1705118 |
| _version_ | 1797678392804376576 |
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| author | Hamze Beati Alistair Langlands Sara ten Have H.-Arno J. Müller |
| author_facet | Hamze Beati Alistair Langlands Sara ten Have H.-Arno J. Müller |
| author_sort | Hamze Beati |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-11T22:59:03Z |
| format | Article |
| id | doaj.art-ed9f1781c79142c6bac0bb4f1cc889d9 |
| institution | Directory Open Access Journal |
| issn | 1933-6934 1933-6942 |
| language | English |
| last_indexed | 2024-03-11T22:59:03Z |
| publishDate | 2020-10-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Fly |
| spelling | doaj.art-ed9f1781c79142c6bac0bb4f1cc889d92023-09-21T15:09:08ZengTaylor & Francis GroupFly1933-69341933-69422020-10-01141-4102810.1080/19336934.2019.17051181705118SILAC-based quantitative proteomic analysis of Drosophila gastrula stage embryos mutant for fibroblast growth factor signallingHamze Beati0Alistair Langlands1Sara ten Have2H.-Arno J. Müller3University of KasselUniversity of DundeeUniversity of DundeeUniversity of KasselQuantitative 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.http://dx.doi.org/10.1080/19336934.2019.1705118drosophilasilacfibroblast growth factorcell signallingproteomics |
| spellingShingle | Hamze Beati Alistair Langlands Sara ten Have H.-Arno J. Müller SILAC-based quantitative proteomic analysis of Drosophila gastrula stage embryos mutant for fibroblast growth factor signalling Fly drosophila silac fibroblast growth factor cell signalling proteomics |
| title | SILAC-based quantitative proteomic analysis of Drosophila gastrula stage embryos mutant for fibroblast growth factor signalling |
| title_full | SILAC-based quantitative proteomic analysis of Drosophila gastrula stage embryos mutant for fibroblast growth factor signalling |
| title_fullStr | SILAC-based quantitative proteomic analysis of Drosophila gastrula stage embryos mutant for fibroblast growth factor signalling |
| title_full_unstemmed | SILAC-based quantitative proteomic analysis of Drosophila gastrula stage embryos mutant for fibroblast growth factor signalling |
| title_short | SILAC-based quantitative proteomic analysis of Drosophila gastrula stage embryos mutant for fibroblast growth factor signalling |
| title_sort | silac based quantitative proteomic analysis of drosophila gastrula stage embryos mutant for fibroblast growth factor signalling |
| topic | drosophila silac fibroblast growth factor cell signalling proteomics |
| url | http://dx.doi.org/10.1080/19336934.2019.1705118 |
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