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

项目摘要 与疾病状态相关的遗传和表观遗传变异的测序显示，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模板化过程生成可访问性的重塑剂。尽管这些复合体具有良好的- 在许多情况下，BAF-家族复合物的机制是由特定的调节作用， 信号仍然知之甚少。此外，整个基因组中的可接近位点以极端的 这些复合物的异质性重塑，但这种异质性的基础， 重塑特异性，以及启动后对转录的影响在很大程度上仍然未知。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， 正常和疾病状态。为了应对这一挑战，我们将开发和联合收割机在表观遗传学的新工具， 化学生物学和显微镜。我们将利用快速的技术来操纵生物体内的BAF活性 细胞使用细胞渗透性分子，并使用敏感，无偏见的方法测量结果， 包括表观基因组学和活细胞显微镜。我们将使用这些工具来回答以下基本问题： ATP依赖的染色质重塑特异性，包括：（1）BAF（SWI/SNF）复合物如何调节 通过手机信号吗(2)为什么不同部位对染色质重塑的反应不同？(3)染色质是如何 启动后重塑影响转录？揭示了这些人使用的基本机制 这些因素有很大的希望，使强大的干预战略，对各种人类疾病。

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.