Multi-Scale Genetics of Transcription Network: Understanding the Regulatory Roles of All 300 Transcription Factors from a Single Organism Escherichia coli

A. Ishihama, H. Ogasawara, T. Shimada, J. Teramoto, A. Hasegawa, Y. Umezawa, K. Yabuki, Y. Ishida, T. Inaba, A. Kori, K. Yamada, Y. Kitai, N. Kobayashi, D. Kato, K. Yamamoto
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Abstract

The modulation of gene selectivity of RNA polymerase is the key step in the bacterial response to environmental changes. The gene selectivity of RNA polymerase is modulated after interaction with seven species of sigma factor, the promoter recognition subunit, and a total of about 300 DNA-binding transcription factors in the second step. In order to get insights into the molecular mechanisms underlying the global regulation of genome expression, our effort has been focused on the identification of genes and promoters under the control of each transcription factor. For this purpose, we have developed the 'Genomic SELEX' system, which allows the quick identification of DNA sequences recognized in vitro by transcription factors. Regulation in vivo of the target genes predicted after the 'Genomic SELEX' was examined by analysis of individual transcripts in the presence and absence of the test transcription factors. The regulated promoter activity was also examined using the newly constructed two-fluorescent-protein promoter assay vector pGRP. Results altogether indicate the involvement of complex networks between transcription factors and promoters in the global regulation of genome transcription.
转录网络的多尺度遗传学:了解来自单一有机体大肠杆菌的所有300个转录因子的调节作用
RNA聚合酶基因选择性的调控是细菌对环境变化反应的关键步骤。在第二步中,RNA聚合酶与7种sigma因子、启动子识别亚基以及总共约300种dna结合转录因子相互作用,调控基因的选择性。为了深入了解基因组表达全局调控的分子机制,我们的工作一直集中在鉴定每个转录因子控制下的基因和启动子。为此,我们开发了“基因组SELEX”系统,该系统可以快速鉴定体外转录因子识别的DNA序列。通过分析存在和不存在测试转录因子的个体转录本,对“基因组SELEX”预测后的靶基因在体内的调节进行了检查。用新构建的双荧光蛋白启动子检测载体pGRP检测调控启动子活性。结果表明,转录因子和启动子之间的复杂网络参与了基因组转录的全局调控。
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