Long-range nucleosome mapping of the HOX gene cluster

描述(由申请人提供)： 核过程(即转录、复制和修复)需要特定的DNA调控序列对反式作用因子可及。目前由染色质结构变化介导的基因调控模型建立在许多单个基因座的研究结果基础上。这项建议旨在开发模型来协调远距离多个基因的高阶染色质结构的变化。实验室开发的核小体间切割和检测方案的高通量自动化将使我们能够研究多个基因的远程核小体定位。生物信息学将被用于绘制核小体位置图，并可能导致识别新的调控位点。将研究HOX基因簇，其中差异基因表达对于造血分化是必不可少的。多梳蛋白和三胸蛋白直接参与HOX基因簇，生化和遗传分析将用于研究这些染色质重构体如何调节核小体定位以调节HOX基因的表达。所提出的方法将使我们能够更好地理解复杂的遗传事件如何通过高阶染色质结构的变化来调节。

DESCRIPTION (provided by applicant): Nuclear processes (i.e. transcription, replication, and repair) require the accessibility of specific DNA regulatory sequences to trans-acting factors. Current models of gene regulation mediated by changes in chromatin structure have been built upon the findings of many individual loci. This proposal aims to develop models for coordinating changes in higher-order chromatin structure for multiple genes over long distances. High throughput automation of an inter-nucleosomal cleavage-and-detection protocol developed in the lab will enable us to investigate long-range nucleosome positioning for multiple genes. Bioinformatics will be used to map nucleosome positions and may lead to the identification of novel regulatory sites. The HOX gene clusters, where differential gene expression is essential for hematopoietic differentiation, will be studied. The Polycomb group and trithorax group proteins are directly involved at the HOX gene clusters and biochemical and genetic assays will be used to examine how these chromatin remodelers regulate nucleosome positioning to mediate HOX gene expression. The proposed methodologies will allow us to better understand how complex genetic events can be regulated by changes in higher-order chromatin structure.