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的可及性。目前的染色质结构变化介导的基因调控模型是建立在许多单个基因座的发现基础上的。该提案旨在开发用于协调多个基因在长距离上的高阶染色质结构变化的模型。 在实验室中开发的核小体间切割和检测协议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。生物信息学将被用来定位核小体的位置，并可能导致新的监管网站的识别。HOX基因簇，其中差异基因表达是造血分化所必需的，将进行研究。Polycomb组和trithorax组蛋白直接参与HOX基因簇，生物化学和遗传测定将用于研究这些染色质重塑物如何调节核小体定位以介导HOX基因表达。所提出的方法将使我们能够更好地了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.