SILAC-based quantitative proteomic analysis of Drosophila gastrula stage embryos mutant for fibroblast growth factor signalling.

IF 2.4 4区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Fly Pub Date : 2020-03-01 Epub Date: 2019-12-24 DOI:10.1080/19336934.2019.1705118

Hamze Beati, Alistair Langlands, Sara Ten Have, H-Arno J Müller

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引用次数: 0

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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基于silac的果蝇原肠期胚胎突变体成纤维细胞生长因子信号的定量蛋白质组学分析。

定量蛋白质组学分析结合遗传学为发育细胞信号研究提供了有力的工具。黑腹果蝇是研究发育和疾病最广泛使用的遗传模型之一。本研究将定量蛋白质组学与遗传选择相结合，以确定原肠期果蝇胚胎中无情(Htl)成纤维细胞生长因子(FGF)受体信号缺失后蛋白质组的变化。我们提出了一个强大的，单代SILAC(稳定同位素标记与氨基酸在细胞培养)方案标记蛋白质在早期胚胎。为了选择原肠胚前阶段的纯合突变胚，我们开发了一个独立的遗传标记。我们的分析检测了htl突变胚胎在原肠胚形成过程中整体蛋白质组的定量变化。我们确定了不同种类的下调和上调蛋白，网络分析表明了每一类蛋白的功能相关组。此外，我们还发现了磷酸肽丰度的变化。总之，我们的定量蛋白质组学分析揭示了htl突变胚胎中代谢、核质、细胞骨架和运输蛋白的整体变化。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Fly 生物-生化与分子生物学

CiteScore

2.90

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： Fly is the first international peer-reviewed journal to focus on Drosophila research. Fly covers a broad range of biological sub-disciplines, ranging from developmental biology and organogenesis to sensory neurobiology, circadian rhythm and learning and memory, to sex determination, evolutionary biology and speciation. We strive to become the “to go” resource for every researcher working with Drosophila by providing a forum where the specific interests of the Drosophila community can be discussed. With the advance of molecular technologies that enable researchers to manipulate genes and their functions in many other organisms, Fly is now also publishing papers that use other insect model systems used to investigate important biological questions. Fly offers a variety of papers, including Original Research Articles, Methods and Technical Advances, Brief Communications, Reviews and Meeting Reports. In addition, Fly also features two unconventional types of contributions, Counterpoints and Extra View articles. Counterpoints are opinion pieces that critically discuss controversial papers questioning current paradigms, whether justified or not. Extra View articles, which generally are solicited by Fly editors, provide authors of important forthcoming papers published elsewhere an opportunity to expand on their original findings and discuss the broader impact of their discovery. Extra View authors are strongly encouraged to complement their published observations with additional data not included in the original paper or acquired subsequently.