SWING LIGAND-REGULATED ENHANCER-DEPENDENT TRANSCRIPTION

摆动配体调节的增强子依赖性转录

• 批准号: 10446924
• 负责人: Dario Meluzzi
• 金额: $ 11.85万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-04 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10446924
• 关键词: Acute; Architecture; Award; Binding; Biological Assay; Chromosomes; Complement; Complex; Cytoplasmic Granules; DNA; DNA Damage; DNA Repair; Data; Enhancers; Estrogen Receptor alpha; Estrogen Receptors; Estrogens; Event; G22P1 gene; Gene Activation; Genes; Genetic Transcription; Genomic approach; Genomics; Grant; Heterochromatin; Housekeeping Gene; Informatics; Knowledge; Licensing; Ligands; Link; Location; Maps; Mediating; Molecular; National Institute of Diabetes and Digestive and Kidney Diseases; Nuclear; Outcome; Parents; Phase; Positioning Attribute; Property; Proteins; Proteomics; RNA; Research Personnel; Resolution; Role; Shapes; Signal Transduction; Single-Stranded DNA; Structure; Testing; Time; Transcription Elongation; Transcriptional Regulation; Type I DNA Topoisomerases; base; career; cellular imaging; experimental study; genome-wide; insight; novel strategies; parent grant; programs; promoter; protein complex; recruit; response

ABSTRACT In response to estrogen-dependent gene activation, the rapid assembly of the MegaTrans complex at responsive enhancers, is suggested to assemble an RNP condensate with phase separation-like properties. The resultant condensate is required for formation of homotypic cooperative enhancer networks, and initial data suggest that these activated enhancers become rapidly associated with specific sub-nuclear bodies. Further, the enhanced transcription of enhancers located separated by many TADs by linear distance may come into closer proximity response to E2. Here, we hypothesize that by linking new mechanistic principles of ERα-bound enhancer, promoter activation events to roles of transcription at chromosomal boundaries in chromosomal architectures and relocation into specific subnuclear architectural structures, can provide mechanistic insights into the activation of large regulated enhancer programs. First, we will explore a new approach to detect single strand DNA nicking genome-wide to uncover a previously-overlooked, but required, mechanism for ligand- and signal-dependent acute activation of enhancer programs. This would uncover a component of the DNA damage program that here instead serves as required coactivators for regulated enhancer activation. We will test the hypothesis that the strongest chromosomal boundaries harbor universally-expressed genes, and that they interact both intra- and inter- chromosomally, impacting chromosomal architecture. This supports a functional association of active chromosomal boundaries with specific subnuclear architectural structures. This R03 award would permit me to generate the essential data required to establish new principles regarding dynamic enhancer activation programs and interactions with specific subnuclear architectural structures for ligand-regulated gene transcriptional programs, providing a strong experimental basis for subsequent submission of an R01 grant.

摘要 作为对雌激素依赖基因激活的反应，MegaTrans复合体的快速组装 建议使用增强剂来组装具有类似相分离性质的RNP凝析油。由此产生的 凝析油是形成同型协同增强子网络所必需的，初步数据表明 这些激活的增强子迅速与特定的亚核小体联系在一起。此外，增强的 被许多TADs直线距离隔开的增强子的转录可能更接近 对雌二醇的反应。在这里，我们假设通过将ERα结合的增强子、启动子的新机制原理联系起来 激活事件对染色体结构中染色体边界转录的作用和重新定位 特定的亚核结构，可以为大分子调控的激活提供机械性的见解 增强程序。首先，我们将探索一种新的方法来检测全基因组范围内的单链DNA划痕 揭示一种先前被忽视但必需的配体和信号依赖的急性激活机制 增强程序。这将揭示DNA损伤计划的一个组成部分，这里的作用是 受调控的增强子激活所需的辅助激活剂。我们将检验这一假设，最强的 染色体边界包含普遍表达的基因，并且它们在细胞内和细胞间相互作用。 在染色体上，影响着染色体的结构。这支持Active的功能关联 具有特定亚核结构的染色体边界。这个R03奖将允许我 生成建立有关动态增强剂激活计划的新原则所需的基本数据 以及与配体调节基因转录的特定亚核结构的相互作用 计划，为随后提交R01赠款提供了坚实的实验基础。