Dimensions in Endocrine Disruption: Altering Receptor Coregulator Interactions

该项目旨在确定可能的环境内分泌干扰物通过一种不寻常的机制发挥作用。 在配体与核激素受体相互作用后起作用的，即通过直接抑制 或增强核激素受体/辅活化子相互作用本身。这个目标一定会实现的 通过开发时间分辨荧光分析，以强健和可靠的方式探测相互作用 带有辅助激活剂的核受体。这些检测方法适用于高通量筛选，以及 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，以确定可能的辅活化子结合抑制物 (CBIS)或可能存在于环境中的共激活剂结合电位剂(CBPS)。后置复合词 已被确定为可能的内分泌功能干扰物或增强剂，它们将受到 二次化验以确认它们确实通过这些不寻常的CBI或CBP机制起作用。这些 检测也将使用荧光共振能量转移方法，但要经过精心挑选 整体成分浓度的变化以阐明其作用机制。最后，基于细胞的分析 例如哺乳动物-2杂交、共转染报告基因分析或激素反应监测 基因产品将更明确地展示可能产生的生物或生理效应 暴露在这些化合物中。这个项目应该有助于精确定位一个新的内分泌干扰部位 核受体信号通路，并确定在这一水平上工作的化合物。这也应该会有帮助 确定环境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，这是由于中断 或增强核受体调节的信号转导，该信号转导作用于配体-受体后 交互级别。最终，这个项目应该有助于识别存在于我们的 环境保护，并有望指导采取措施，最大限度地减少公众对这些物质的暴露。 内分泌干扰物是干扰内分泌系统的外源性化合物，改变荷尔蒙 动作和它在全身传递的信息。该项目旨在确定可能的内分泌。 通过一种不同寻常的机制起作用的破坏者。将对大量化合物进行筛选，以确定 通过这一不寻常的过程抑制或增强内分泌作用的那些细胞的结构特征。有了这个 根据信息，可以采取措施尽量减少人类对这些新型内分泌干扰物的接触。

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.