Grainger David C, Hurd Douglas, Harrison Marcus, Holdstock Jolyon, Busby Stephen J W
School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom.
Proc Natl Acad Sci U S A. 2005 Dec 6;102(49):17693-8. doi: 10.1073/pnas.0506687102. Epub 2005 Nov 21.
Chromatin immunoprecipitation and high-density microarrays have been used to monitor the distribution of the global transcription regulator Escherichia coli cAMP-receptor protein (CRP) and RNA polymerase along the E. coli chromosome. Our results identify targets occupied by CRP and genes transcribed by RNA polymerase in vivo. Many of the loci of CRP binding are at known CRP regulated promoters. However, our results show that CRP also interacts with thousands of weaker sites across the whole chromosome and that this "background" binding can be used as a probe for organization within the E. coli folded chromosome. In rapidly growing cells, we show that the major sites of RNA polymerase binding are approximately 90 transcription units that include genes needed for protein synthesis. Upon the addition of rifampicin, RNA polymerase is distributed among >500 functional promoters. We show that the chromatin immunoprecipitation and high-density-microarrays methodology can be used to study the redistribution of RNA polymerase induced by environmental stress, revealing previously uncharacterized aspects of RNA polymerase behavior and providing an alternative to the "transcriptomics" approach for studying global transcription patterns.
染色质免疫沉淀和高密度微阵列已被用于监测全局转录调节因子大肠杆菌环磷酸腺苷受体蛋白(CRP)和RNA聚合酶沿大肠杆菌染色体的分布。我们的结果确定了体内被CRP占据的靶点以及由RNA聚合酶转录的基因。许多CRP结合位点位于已知的CRP调控启动子处。然而,我们的结果表明,CRP还与整个染色体上数千个较弱的位点相互作用,并且这种“背景”结合可作为探测大肠杆菌折叠染色体内组织的探针。在快速生长的细胞中,我们表明RNA聚合酶结合的主要位点约为90个转录单位,其中包括蛋白质合成所需的基因。加入利福平后,RNA聚合酶分布在500多个功能性启动子之间。我们表明,染色质免疫沉淀和高密度微阵列方法可用于研究环境应激诱导的RNA聚合酶的重新分布,揭示RNA聚合酶行为以前未被表征的方面,并为研究全局转录模式的“转录组学”方法提供了一种替代方法。