Dimensions in Endocrine Disruption: Altering Receptor Coregulator Interactions

本项目旨在确定可能的环境内分泌干扰物通过不寻常的机制工作 在配体与核激素受体相互作用后起作用，即通过直接抑制 或核激素受体/辅激活因子相互作用本身的增强。这一目标将得以实现 通过开发时间分辨荧光分析，以稳健和可靠的方式探测相互作用， 核受体与辅激活剂的关系。这些测定适用于高通量筛选，并且 将在埃默里大学分子图书馆筛选中心使用，这是NIH资助的路线图 我们已获得访问权限的研究资源，以识别可能的辅激活因子结合抑制剂 在一些实施方案中，所述化合物是可能存在于环境中的CBIs或共激活剂结合增强剂（CBP）。化合物后 已被确定为内分泌功能的可能干扰物或增强物，它们将受到 第二次试验，以确认他们确实通过这些不寻常的CBI或CBP机制。这些 测定也将使用荧光共振能量转移方法，但仔细选择 整体组分浓度的改变以阐明机制。最后，基于细胞的分析 例如哺乳动物-2-杂交、共转染报告基因测定或监测对细胞凋亡应答 基因产物将更明确地证明可能的生物学或生理学效应， 暴露在这些化合物中。这个项目应该有助于确定一个新的内分泌干扰网站内 核受体信号通路，并确定在这一水平工作的化合物。它也应该有帮助 确定环境中或常用产品中可能存在的CBI或CBP， 通过一种不寻常的机制导致基因组转录模式的改变， 或增强核受体调节的信号转导，在后配体-受体 互动水平。最终，这个项目应该有助于确定我们体内存在的有害化合物。 环境，并希望指导采取措施，尽量减少公众接触这些物质。 内分泌干扰物是干扰内分泌系统的外源性化合物， 动作和它在身体中传递的信息。该项目旨在确定可能的内分泌 通过一种不寻常的机制起作用的破坏者。大量的化合物将被筛选， 通过这种不寻常的过程抑制或增强内分泌作用的结构特征。与此 如果有足够的信息，可以采取措施尽量减少人类接触这些新型内分泌干扰物。

This project aims to identify possible environmental endocrine disrupters working by an unusual mechanism that operates after the interaction of ligands with nuclear hormone receptors, namely, by the direct inhibition or potentiation of the nuclear hormone receptor/coactivator interaction itself. This goal will be accomplished by developing time-resolved fluorescence assays that probe in a robust and reliable manner the interaction of the nuclear receptors with coactivators. These assays are adaptable to high-throughput screening, and will be used at the Emory University Molecular Libraries Screening Center, an NIH-funded Roadmap Research Resource to which we have obtained access, to identify possible coactivator binding inhibitors (CBIs) or coactivator binding potentiators (CBPs) that might be present in the environment. After compounds have been identified as possible disrupters or potentiators of endocrine function, they will be subject to secondary assays to confirm that they indeed act through these unusual CBI or CBP mechanisms. These assays will also use fluorescence resonance energy transfer methodology, but with carefully selected alterations in the concentrations of integral components to elucidate mechanism. Finally, cell-based assays such as mammalian-2-hybrid, cotransfection reporter gene assays, or monitoring of hormone-responsive gene products will more definitively demonstrate the possible biological or physiological effects arising from exposure to these compounds. This project should help pinpoint a novel endocrine disruption site within the nuclear receptor signaling pathway and identify the compounds that work at this level. It should also help identify possible CBIs or CBPs present in the environment or in commonly used products that could be causing alterations in the pattern of genome transcription by an unusual mechanism, due to the interruption or the potentiation of nuclear receptor-regulated signal transduction that operates at a post ligand-receptor interaction level. Ultimately, this project should help in identifying harmful compounds present within our environment and, hopefully, guide measures to minimize exposure of the public to these substances. Endocrine disrupters are exogenous compounds that interfere with the endocrine system, altering hormone action and the messages it sends throughout the body. This project aims to identify possible endocrine disrupters that act by an unusual mechanism. Large numbers of compounds will be screened to determine structural featurs of those that inhibit or potentiate endocrine action by this unusual process. With this information, steps could be taken to minimize human exposure to these novel types of endocrine disrupters.