A visual screen for diet-regulated proteins in the Drosophila ovary using GFP protein trap lines
A visual screen for diet-regulated proteins in the Drosophila ovary using GFP protein trap lines
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DOI：
10.1016/j.gep.2017.01.001
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发表时间：
2017-01-01
影响因子：
1.2
通讯作者：
Drummond-Barbosa, Daniela
中科院分区：
文献类型：
--
作者：
Hsu, Hwei-Jan;Drummond-Barbosa, Daniela
饮食对繁殖的影响在大量的生物体中得到了很好的证明;然而，关于这种联系的分子机制还有很多东西有待了解。果蝇卵巢对饮食有一个很好的描述，快速和基本可逆的反应。卵巢干细胞及其后代在富含酵母的饮食中比在无酵母（差）饮食中增殖和生长更快，并且早期生殖细胞包囊的死亡，早期卵黄发生卵泡的退化和排卵的部分阻滞进一步导致在差饮食中观察到的产卵减少60倍。多种饮食依赖性因子，包括胰岛素样肽、类固醇蜕皮激素、营养传感器雷帕霉素靶标、AMP依赖性激酶和脂肪细胞因子介导这种复杂的反应。在这里，我们描述了一个视觉屏幕的结果，使用收集的绿色荧光蛋白（GFP）蛋白陷阱线，以确定其他因素可能参与这种反应。在每个GFP蛋白捕获线中，人工GFP外显子与内源蛋白在读框内融合，使得GFP融合模式与相应天然蛋白的水平和亚细胞定位平行。我们鉴定了53种GFP标记的蛋白质，这些蛋白质在将雌性从丰富饮食切换到贫穷饮食后12-16小时在卵巢中表现出水平和/或亚细胞定位的变化，这表明它们是未来功能研究的潜在候选者。(C)2017爱思唯尔B. V.保留所有权利。
The effect of diet on reproduction is well documented in a large number of organisms; however, much remains to be learned about the molecular mechanisms underlying this connection. The Drosophila ovary has a well described, fast and largely reversible response to diet. Ovarian stem cells and their progeny proliferate and grow faster on a yeast-rich diet than on a yeast-free (poor) diet, and death of early germline cysts, degeneration of early vitellogenic follicles and partial block in ovulation further contribute to the similar to 60-fold decrease in egg laying observed on a poor diet. Multiple diet-dependent factors, including insulin-like peptides, the steroid ecdysone, the nutrient sensor Target of Rapamycin, AMP-dependent kinase, and adipocyte factors mediate this complex response. Here, we describe the results of a visual screen using a collection of green fluorescent protein (GFP) protein trap lines to identify additional factors potentially involved in this response. In each GFP protein trap line, an artificial GFP exon is fused in frame to an endogenous protein, such that the GFP fusion pattern parallels the levels and subcellular localization of the corresponding native protein. We identified 53 GFP-tagged proteins that exhibit changes in levels and/or subcellular localization in the ovary at 12-16 hours after switching females from rich to poor diets, suggesting them as potential candidates for future functional studies. (C) 2017 Elsevier B.V. All rights reserved.