Determinants of genome-wide activity and specificity of SWI/SNF family chromatin remodeling

项目摘要 对与疾病状态相关的遗传和表观遗传变异的测序表明，ATP- 依赖的染色质重构体是许多疾病中最常被破坏的基因之一。在……里面 哺乳动物、BAF(SWI/SNF)、PBAF和GBAF复合体是高度保守的依赖于ATP的染色质 为DNA模板化过程生成可访问性的重建器。尽管这些建筑群有很好的- 在许多情况下记录的角色，BAF-家族复合体受特定调控的机制 人们仍然对信号知之甚少。此外，整个基因组中可访问的位置对极端 异质性对这些复合体的重塑，然而这种异质性的基础，物理起源 重塑的特异性以及启动后对转录的影响在很大程度上仍不清楚。改进 对这些原理的理解将为操纵基因表达提供强有力的机会。 正常状态和疾病状态。为了应对这一挑战，我们将开发和结合表观遗传学的新工具， 化学生物学和显微镜。我们将利用快速技术来控制生活中的BAF活动 使用细胞可渗透分子的细胞，并使用敏感、公正的方法测量结果， 包括表观基因组学和活细胞显微镜。我们将使用这些工具回答以下基本问题 依赖于ATP的染色质重塑特异性，包括：(1)BAF(SWI/SNF)复合体是如何调节的 通过细胞信号？(2)为什么部位对染色质重塑有不同的反应？(3)染色质是如何 重塑对启动后转录的影响？揭示了这些人使用的基本机制 因素很有希望实现对各种人类疾病的强有力的干预策略。

Project Summary Sequencing of genetic and epigenetic variants associated with disease states has revealed that ATP- dependent chromatin remodelers are among the most frequently disrupted genes in a number of diseases. In mammals, BAF (SWI/SNF), PBAF, and GBAF complexes are highly conserved ATP-dependent chromatin remodelers that generate accessibility for DNA-templated processes. Although these complexes have well- documented roles in many contexts, the mechanisms by which BAF-family complexes are regulated by specific signals remains poorly understood. Additionally, accessible sites across the genome respond with extreme heterogeneity to remodeling by these complexes, yet the basis of this heterogeneity, the physical origins of remodeling specificity, as well as the effects on transcription after initiation remain largely unknown. Improved understanding of these principles would provide powerful opportunities to manipulate gene expression in normal and disease states. To address this challenge, we will develop and combine new tools in epigenetics, chemical biology, and microscopy. We will make use of rapid technologies to manipulate BAF activity in living cells using cell-permeable molecules, and measure the outcomes using sensitive, unbiased approaches, including epigenomics and live-cell microscopy. We will use these tools to answer essential questions about ATP-dependent chromatin remodeling specificity, including: (1) How are BAF (SWI/SNF) complexes regulated via cellular signals? (2) Why do sites respond differently to chromatin remodeling? (3) How does chromatin remodeling influence transcription after initiation? Revealing the fundamental mechanisms used by these factors holds great promise to enable powerful intervention strategies for diverse human diseases.