Promoter Regulation in Response to Environmental Stress

描述（由申请人提供）：真核基因的调控涉及数百种蛋白质。它们如何在组成基因组的数千个基因中共同工作尚不清楚。当细胞基因组响应于环境压力等信号事件而重新编程时，这种相互作用是如何变化的，人们对此知之甚少。 这里提出的工作是一个项目的继续，广泛调查转录调控蛋白位于整个基因组的模式真核生物酿酒酵母，以及它们易位时，基因组被环境信号，如热休克和其他压力重新编程。热冲击为单元提供快速且简单的编程事件。 目前对该项目的研究已经揭示了新的见解，基因调控表明，前起始复合物部分组装在核小体启动子区域。然而，完全组装需要去除位于启动子上游侧的“-1”核小体。 在正常和热休克条件下，通过ChIP芯片以低分辨率确定了~200个基因和染色质调节蛋白的基因组位置。目前正在分析数据。将通过为ChIP-seq改造ChIP-chip文库来实现高分辨率映射。甚至更高的分辨率映射将实现对一组选定的因素使用试剂来修剪的ChIP DNA。结合事件之间的关系将提供新的见解转录复合体的组装和调控。

DESCRIPTION (provided by applicant): The regulation of eukaryotic genes involves many hundreds of proteins. How they work together at the many thousands of genes that comprise a genome is not known. Little is known about how this interplay changes as cellular genomes are reprogrammed in response to signaling events such as environmental stress. The work proposed here is a continuation of a project to broadly survey where transcriptional regulatory proteins are located throughout the genome of the model eukaryotic organism Saccharomcyes cerevisiae, and where they translocate to when the genome is reprogrammed by environmental signals, such as heat shock and other stresses. Heat shock provides a rapid and simple programming event for the cell. Current studies on this project have already revealed novel insights into gene regulation by demonstrating that pre-initiation complexes partially assemble in nucleosome-free promoter regions. However, full assembly requires removal of the "-1" nucleosome located on the upstream side of the promoter. The genomic location of ~200 gene- and chromatin-regulatory proteins have been determined at low resolution by ChIP-chip, under normal and heat shock conditions. The data is now being analyzed. High resolution mapping will be achieved by retrofitting the ChIP-chip libraries for ChIP-seq. Even higher resolution mapping will be achieved on a select set of factors using reagents to trim the ChIP DNA. Relationships among binding events will provide new insights into transcription complex assembly and regulation.