Dynamic regulation of Drosophila nuclear receptor activity in vivo
Dynamic regulation of Drosophila nuclear receptor activity in vivo
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DOI：
10.1242/dev.02512
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发表时间：
2006-09-15
期刊：
影响因子：
4.6
通讯作者：
Krause, Henry M.
中科院分区：
文献类型：
--
作者：
Palanker, Laura;Necakov, Aleksandar S.;Krause, Henry M.
核受体是一个转录因子大家族，在发育、变态、代谢和疾病中发挥重要作用。为了确定核受体如何、在何处以及何时受到小型化学配体和/或蛋白质伴侣的调节，我们使用“配体传感器”系统来可视化活体发育动物中 18 种果蝇核受体中每一种的空间活动模式。建立了表达与酵母 GAL4 的 DNA 结合结构域融合的每个核受体的配体结合结构域的转基因系。当与 GAL4 响应报告基因结合时，融合蛋白显示出组织和阶段特异性的激活模式。我们表明，这些反应准确地反映了内源性和外源性添加激素的存在，并且它们可以通过核受体伴侣蛋白进行调节。羊膜浆膜、卵黄、中肠和脂肪体在脂质储存、代谢和发育时间中起重要作用，被认为是核受体活动的常见部位。我们还看到激活的动态变化，这表明配体和/或辅因子产生的彻底变化。使用该系统筛选小型化合物库，鉴定出角补骨脂素当归素和昆虫生长调节剂苯氧威作为 Ultraspiracle (USP) 配体结合域的激活剂。这些结果证明了该系统对于核受体途径的功能剖析以及用于开发新的受体激动剂和拮抗剂的实用性，这些受体激动剂和拮抗剂可用于调节代谢和疾病以及开发更有效的昆虫控制手段。
Nuclear receptors are a large family of transcription factors that play major roles in development, metamorphosis, metabolism and disease. To determine how, where and when nuclear receptors are regulated by small chemical ligands and/or protein partners, we have used a 'ligand sensor' system to visualize spatial activity patterns for each of the 18 Drosophila nuclear receptors in live developing animals. Transgenic lines were established that express the ligand binding domain of each nuclear receptor fused to the DNA-binding domain of yeast GAL4. When combined with a GAL4-responsive reporter gene, the fusion proteins show tissue- and stage-specific patterns of activation. We show that these responses accurately reflect the presence of endogenous and exogenously added hormone, and that they can be modulated by nuclear receptor partner proteins. The amnioserosa, yolk, midgut and fat body, which play major roles in lipid storage, metabolism and developmental timing, were identified as frequent sites of nuclear receptor activity. We also see dynamic changes in activation that are indicative of sweeping changes in ligand and/or co-factor production. The screening of a small compound library using this system identified the angular psoralen angelicin and the insect growth regulator fenoxycarb as activators of the Ultraspiracle (USP) ligand-binding domain. These results demonstrate the utility of this system for the functional dissection of nuclear receptor pathways and for the development of new receptor agonists and antagonists that can be used to modulate metabolism and disease and to develop more effective means of insect control.