SWING LIGAND-REGULATED ENHANCER-DEPENDENT TRANSCRIPTION

摆动配体调节的增强子依赖性转录

• 批准号: 10599977
• 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/10599977
• 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; Grant; Heterochromatin; Housekeeping Gene; Informatics; Knowledge; Licensing; Ligands; Link; Maps; Mediating; Molecular; National Institute of Diabetes and Digestive and Kidney Diseases; Outcome; Parents; Phase; Physical condensation; 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; career; cellular imaging; experimental study; genome-wide; genomic locus; 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 在雌激素依赖性基因激活的反应中，MegaTrans复合物的快速组装在响应性细胞中发生。 增强剂，建议组装具有类似相分离性质的RNP缩合物。所得 缩合物是形成同型合作增强子网络所必需的，并且初步数据表明， 这些活化的增强子迅速与特定的亚核体结合。此外，增强 被许多TADs以线性距离分开的增强子的转录可能变得更接近 回复E2在这里，我们假设通过连接ERα结合增强子，启动子 激活事件对染色体结构中染色体边界处转录的作用以及重新定位到 特定的亚核架构结构，可以提供机制的见解激活大的调节 增强程序。首先，我们将探索一种新的方法来检测单链DNA切口全基因组， 揭示了一个以前被忽视的，但需要的，配体和信号依赖性急性激活的机制， 增强程序。这将揭示DNA损伤程序的一个组成部分，在这里， 调节增强子激活所需的辅激活剂。我们将检验一个假设， 染色体边界窝藏普遍表达的基因，它们相互作用的内部和之间， 影响染色体结构。这支持活动的 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子激活程序的新原则所需的基本数据 与配体调控基因转录的特定亚核结构的相互作用 计划，为随后提交R01赠款提供了强有力的实验基础。