Nuclear Architecture, NcRNAs and Epigenetics in Transcriptional Regulation by ER

ER 转录调控中的核结构、NcRNA 和表观遗传学

基本信息

批准号：
8708062
负责人：
Chunru Lin
金额：
$ 24.9万
依托单位：
UNIVERSITY OF TX MD ANDERSON CAN CTR
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-04-29 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8708062
关键词：
Address Affect Antibodies Architecture Award Binding Biological Assay Breast Cancer Cell Cell Line ChIP-seq Chromatin Complex Cytoplasmic Granules Data Databases Development Disease Environment Epigenetic Process Epithelial Cells Estradiol Estrogen Receptor alpha Estrogen Receptors Estrogens Event Excision Female Functional RNA Gene Activation Gene Expression Gene Expression Profile Gene Expression Regulation Gene Targeting Genes Genetic Transcription Goals Growth Histone Code Histones Homeostasis In Vitro Ligands Location Mammary gland Mediating Metabolic Metabolism Methylation Modification Molecular Nuclear Peptides Play Polycomb Positioning Attribute Process Proteins Reader Reading Regulation Repression Research Personnel Role Signal Transduction Small Interfering RNA Small RNA Structure Tail Testing Therapeutic Time Transcript Transcriptional Regulation base blood glucose regulation cofactor demethylation digital drug development genome-wide glucose metabolism in vivo innovation insight malignant breast neoplasm mammary gland development novel novel strategies preference programs promoter protein function reproductive function response transcription factor transcriptome sequencing

项目摘要

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.

项目概要雌二醇通过雌激素受体 α (ER¿) 发挥作用，对于发育和发育至关重要。女性生殖功能的稳态，对许多转录程序进行关键控制，包括这些调节增殖和代谢的分子机制。调解基因激活中的共激活子/辅阻遏物交换已得到相当好的阐明。 , 染色质特定位点的修饰符和动态定位对于建立精确的转录至关重要然而，目前尚不清楚这些因素是如何发挥作用的。在不同的亚核结构中进行空间调节以及这种调节如何有助于基因调节。我们最近的发现表明信号诱导的转录单元从我将转录抑制室转移到一个宽松的环境中。 (ncRNA) TUG1 和 NEAT2 控制 ER¿目标基因从转录抑制中重新定位多梳体 (PcG) 通过选择性相互作用作用于染色质间颗粒的基因激活环境 ER 上存在甲基化和未甲基化的 Polycomb 2 蛋白 (Pc2)靶基因启动子。为了解决这个长期目标，我建议从三个具体目标来研究假设：1）定义非组蛋白甲基化/去甲基化事件和 ncRNA 作为负责基因组的新型分子策略宽 ER¿转录程序；2) 确定两种 ncRNA TUG1 和 NEAT2 的潜在作用，分别位于 PcG 和染色质间颗粒中，控制 ER¿ 的重新定位靶基因这两个亚核结构之间的差异，取决于 Pc2 甲基化对雌激素的反应状态； 3) 研究 ncRNA 在调节组蛋白密码“读者”能力方面的潜在变构作用，例如 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非甲基化 Pc2 的全基因组位置通过使用特定抗体的 ChIP-Seq 分析以及 Pc2 或其他活性“读取器”的功能作用，以及 ncRNA 控制 ER¿结合 GRO-Seq、RNA-Seq 分析和 siRNA 进行转录组分析我还将研究 ER 的潜在重新定位策略。 -来自PcG的调控基因 E2 刺激后小体与染色质间颗粒的结合及其潜在机制 TUG1 或 NEAT2 ncRNA 指导 Pc2 识别组蛋白尾部修饰的偏好。综上所述，拟议的研究评估了一个新概念，即 ER¿目标基因位点在之间重新定位功能结构上不同的核结构以及几种发挥关键作用的新型调节剂的作用在 ER 中的作用这些发现将为药物开发提供创新目标。基于大量的初步数据，我相信有许多有趣的治疗意义。在获奖期间实现这些目标并成为一名独立研究者。