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

• 批准号: 8913154
• 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
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-29 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8913154
• 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; Fluorescent in Situ Hybridization; 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; Non-Histone Chromosomal Proteins; Nuclear; Peptides; Play; Polycomb; Positioning Attribute; Process; Protein Methylation; Proteins; Reader; Reading; Regulation; Repression; Research Personnel; Role; Signal Transduction; Small Interfering RNA; Small RNA; Structure; Tail; Testing; Therapeutic; Time; Transcript; Transcriptional Regulation; Untranslated RNA; base; blood glucose regulation; cofactor; demethylation; digital; drug development; genome-wide; global run on sequencing; glucose metabolism; histone methylation; 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

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.

项目摘要 雌激素通过雌激素受体α（ER β）发挥作用，对发育和 雌性生殖功能的稳态，对许多转录程序发挥关键控制作用， 包括这些调节增殖和代谢。ER β介导的分子机制 基因激活中的辅激活子/辅阻遏子交换已经得到合理的阐明。ER�，染色质 修饰子和特定基因座的动态定位对于建立精确的转录调控系统都是必不可少的。 调节和实现基因表达的正确模式。不过，目前尚不清楚这些因素如何 在不同的亚核结构的空间调控，以及这种调控如何可能有助于基因调控。 我们最近的研究结果表明，信号诱导的转录单位从一个 转录抑制区室到允许环境。我假设非编码RNA （ncRNA）TUG 1和NEAT 2控制ER β靶基因从转录抑制基因中重新定位， 多梳体（Polycomb bodies，PcGs）与染色质间颗粒的基因激活环境，通过选择性相互作用 甲基化和未甲基化的Polycomb 2蛋白（Pc 2）存在于ER靶基因启动子上。到 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作为一种新的分子策略负责基因组- 广泛的ER β转录程序; 2）确定两种ncRNA，TUG 1和NEAT 2的潜在作用， 分别位于PcGs和染色质间颗粒，控制ER靶基因的迁移 这两个亚核结构之间的差异，取决于雌激素对Pc 2甲基化的反应状态; 3)为了研究ncRNA在调节组蛋白密码“阅读器”能力中的潜在变构作用， 例如Pc 2，以识别组蛋白尾部修饰。 为了实现这些目标，我建议确定甲基化和非甲基化的Pc 2的全基因组位置， 通过使用特异性抗体的ChIP-Seq分析和Pc 2或其他活性“阅读器”的功能作用， 通过GRO-Seq，RNA-Seq分析和siRNA的组合控制ER β转录组中的ncRNA 击倒战略我还将研究雌激素受体β调节基因从PcG中的潜在重新定位 E2刺激后，体与染色质间颗粒的结合及其潜在机制， TUG 1或NEAT 2 ncRNA指导组蛋白尾部修饰的Pc 2识别偏好。 综上所述，这项研究评估了一个新的概念，即ER靶基因位点在 功能上不同的核结构和几种新的调节剂的作用，发挥关键作用， ER靶基因调控的作用。这些发现将为药物开发提供创新靶点， 一些有趣的治疗意义。根据大量的初步数据，我相信 在获奖期间实现这些目标，并成为独立调查员。

项目成果

Poly-ADP ribosylation of PTEN by tankyrases promotes PTEN degradation and tumor growth.

• DOI: 10.1101/gad.251785.114
• 发表时间: 2015-01-15
• 期刊: Genes & development
• 影响因子: 10.5
• 作者: Li N;Zhang Y;Han X;Liang K;Wang J;Feng L;Wang W;Songyang Z;Lin C;Yang L;Yu Y;Chen J
• 通讯作者: Chen J