Nuclear Architecture, NcRNAs and Epigenetics in Transcriptional Regulation by ER

项目摘要 雌二醇通过雌激素受体α(ER�)发挥作用，是发育和发育所必需的。 雌性生殖功能的动态平衡，对许多转录程序施加关键控制， 包括这些调控的增殖和新陈代谢。内质网�介导的分子机制 基因激活中的辅活化子/辅抑制子交换已经被很好地阐明。ER�，染色质 修饰物和特定基因座的动态定位对于建立精确的转录都是必不可少的 调节和实现正确的基因表达模式。然而，目前还不清楚这些因素是如何产生的 在不同的亚核结构中空间调控，以及这种调控如何有助于基因调控。 我们最近的发现证明了信号诱导的转录单位从 转录抑制隔间到允许的环境。我假设非编码的RNA (NcRNAs)TUG1和NEAT2控制ER�靶基因从转录抑制中重新定位 多梳小体(PCGS)通过选择性相互作用对染色质间颗粒的基因激活环境的影响 ER�靶基因启动子上存在甲基化和未甲基化的聚梳2蛋白(PC2)。至 为了解决这一长期目标，我建议从三个具体目标来研究这一假说：1)定义非 组蛋白甲基化/去甲基化事件和ncRNAs作为一种新的分子策略负责基因组... 广泛的ER�转录程序；2)确定两个ncRNATUG1和NEAT2的潜在作用， 分别位于PCGS和染色质间颗粒中，控制ER�靶基因的重新定位 在这两个亚核结构之间，取决于PC2甲基化对雌激素的反应状态； 3)为了研究ncRNA在调节组蛋白密码的“读取器”能力中的潜在变构作用， 如PC2，用于识别组蛋白尾部修饰。 为了实现这些目标，我建议定义甲基化与未甲基化的PC2在全基因组中的位置 通过使用特定抗体和PC2或其他活跃的“读取器”的功能作用和 基因序列、序列分析和小干扰RNA在调控ER�转录组中的作用 击倒战略。我还将研究pcg中ER�调节基因的潜在重新定位 E_2刺激下机体对染色质间颗粒的结合及其潜在机制 TUG1或NEAT2 ncRNAs引导PC2识别组蛋白尾部修饰的偏好。 综上所述，这项研究评估了ER�靶基因座位在 功能上不同的核结构结构和几个发挥关键作用的新的调节器的作用 在ER�靶基因调控中的作用。这些发现将为药物开发提供创新的目标 一些耐人寻味的治疗暗示。根据广泛的初步数据，我有信心 在获奖期间实现这些目标并成为一名独立调查员。

Project Summary Estradiol, which acts through the Estrogen Receptor alpha (ER�), is essential for the development and homeostasis of female reproductive function, exerting critical control of many transcriptional programs, including these regulatory proliferation, and metabolism. The molecular mechanism used by ER� to mediate coactivator/corepressor exchanges in gene activation has been reasonably well elucidated. ER�, chromatin modifiers and dynamic positioning of particular loci are all essential for establishing precise transcriptional regulation and achieving the correct patterns of gene expression. However, it is not clear how these factors are spatially regulated in distinct subnuclear structures and how this regulation may contribute to gene regulation. Our recent findings demonstrated a signal-induced relocation of the transcription units from a transcriptional repressive compartment to a permissive environment. I hypothesize that non-coding RNAs (ncRNAs) TUG1 and NEAT2 control ER� target genes to relocate from the transcriptional repressive Polycomb bodies (PcGs) to the gene activation milieu of the interchromatin granules, by selectively interacting with methylated and unmethylated Polycomb 2 protein (Pc2) present on ER� target gene promoters. To address this long-term goal, I propose to investigate the hypothesis from three specific aims: 1) to define non- histone methylation/demethylation events and ncRNAs as a novel molecular strategy responsible for genome- wide ER� transcriptional programs; 2) to determine the potential role of two ncRNAs, TUG1 and NEAT2, located in PcGs and interchromatin granules, respectively, in controlling relocation of ER� target genes between these two subnuclear structures, depending on the status of Pc2 methylation in response to estrogen; 3) to investigate the potential allosteric role of ncRNA in modulating the ability of "readers" of the histone code, such as Pc2, to recognize histone tail modifications. To achieve these aims, I propose to define the genome-wide location of methylated vs. unmethylated Pc2 by ChIP-Seq analysis using specific antibodies and the functional roles of Pc2 or other active "readers" and ncRNAs in controlling the ER� transcriptome by a combination of GRO-Seq, RNA-Seq analysis and siRNA knockdown strategies. I will also investigate the potential relocalization of ER�-regulated genes from PcG bodies to interchromatin granules upon E2 stimulation and the underlying mechanisms, by which the binding of TUG1 or NEAT2 ncRNAs directs the preference of Pc2 recognition of histone tail modification. Taken together, the proposed study evaluates a new concept that ER� target gene loci relocate between functionally distinct nuclear architectural structures and the roles of several novel modulators that play a critical role in ER� target gene regulation. These findings will provide innovative targets for drugs development for a number of intriguing therapeutic implications. Based on the extensive preliminary data, I am confident of accomplishing these aims in the period of the award and emerging as an Independent Investigator.