Studies of the distribution of Escherichia coli cAMP-receptor protein and RNA polymerase along the E-coli chromosome
Studies of the distribution of Escherichia coli cAMP-receptor protein and RNA polymerase along the E-coli chromosome
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DOI：
10.1073/pnas.0506687102
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发表时间：
2005-12-06
影响因子：
11.1
通讯作者：
Busby, SJW
中科院分区：
文献类型：
--
作者：
Grainger, DC;Hurd, D;Busby, SJW
染色质免疫沉淀和高密度微阵列已被用于监测全球转录调节因子大肠杆菌cAMP受体蛋白（CRP）和RNA聚合酶沿着E. coli染色体。我们的研究结果确定了CRP和RNA聚合酶在体内转录的基因所占据的目标。CRIP结合的许多基因座位于已知的CRIP调节启动子处。然而，我们的研究结果表明，CRP也与整个染色体上数千个较弱的位点相互作用，这种“背景”结合可以用作E. coli折叠染色体。在快速生长的细胞中，我们发现RNA聚合酶结合的主要位点大约有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个功能启动子中。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由环境压力诱导的RNA聚合酶的再分布，揭示了RNA聚合酶行为的以前未表征的方面，并提供了一种替代的“转录组学”的方法来研究全球转录模式。
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 CRIP binding are at known CRIP 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 approximate to 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.