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

项目概要 与疾病状态相关的遗传和表观遗传变异的测序表明 ATP- 依赖性染色质重塑基因是许多疾病中最常被破坏的基因之一。在 在哺乳动物中，BAF (SWI/SNF)、PBAF 和 GBAF 复合物是高度保守的 ATP 依赖性染色质 为 DNA 模板化过程提供可访问性的重塑器。尽管这些复合体具有良好的 记录了在许多情况下的作用，BAF家族复合物受特定调节的机制 信号仍然知之甚少。此外，整个基因组中可访问的位点会做出极端的反应 这些复合物重塑的异质性，但这种异质性的基础， 重塑特异性以及起始后对转录的影响仍然很大程度上未知。改进 了解这些原理将为操纵基因表达提供强大的机会 正常和疾病状态。为了应对这一挑战，我们将开发并结合表观遗传学的新工具， 化学生物学和显微镜学。我们将利用快速技术来操纵生活中的 BAF 活动 使用细胞渗透性分子检测细胞，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检查。我们将使用这些工具来回答以下基本问题 ATP依赖性染色质重塑特异性，包括：（1）BAF（SWI/SNF）复合物如何调控 通过蜂窝信号？ (2) 为什么位点对染色质重塑的反应不同？ (3) 染色质如何 重塑影响启动后的转录？揭示这些所使用的基本机制 因素有望为多种人类疾病提供强有力的干预策略。

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.