Investigation of Comprehensive Vision of Transcriptional regulation in Bacillus subtilis

使用在各种培养条件下生长并暴露于多种环境胁迫的细胞，获得了500多个枯草芽孢杆菌DNA微阵列数据，以进行改变的基因的基因聚类，并使用转录调节突变体，提取转录调节因子的靶基因候选物。通过基因聚类和候选靶基因的鉴定，揭示了一个涉及支链氨基酸生物合成的基本代谢调控网络，包括CcpA、CodY和TnrA的碳、氮调控和RelA的严格调控。我们完成了ECF-sigma因子的全面分子遗传分析，并基本完成了34个双组分调控系统的分子遗传分析。因此，只有几个双组分系统的功能仍然是未知的。此外，我们还对200多个HTH-DNA结合调节蛋白中的马尔R、TetR和MerR家族进行了分子遗传学分析，并对这三个家族的几乎所有蛋白进行了基因芯片分析，确定了至少一个靶基因。特别值得注意的是，我们发现YsiA蛋白参与了全局脂肪酸降解调控。这些五年计划的结果，揭开了B转录网络的大致轮廓。枯草芽孢杆菌，发表在下面列出的39篇科学论文中，以及在科学会议上的100多个演讲。

More than 500 Bacillus subtilis DNA microarray data were obtained, using the cells grown in various cultivation conditions and exposed to numerous environmental stresses, to perform gene clustering of altered genes, and using the transcriptional regulatory mutants, to extract the target gene candidates of the transcriptional regulatory factors. A fundamental network of metabolic regulation concerned with the biosynthesis of branched-chain amino acids involving carbon and nitrogen regulations by CcpA, CodY and TnrA and stringent control conducted by RelA, was revealed from the data of gene clustering and identification of target gene candidates.On the base of the DNA microarray data of DNA-binding transcriptional regulatory factors, we completed comprehensive molecular genetic analysis of ECF-sigma factors, and almost completed that of 34 two-component regulatory systems. Consequently, the function of only several two-component systems remained to be unknown. Moreover, we performed the molecular genetic analysis of the MarR, TetR and MerR families of HTH-DNA binding regulatory proteins among more than 200 proteins, for which the DNA maicroarray analysis of were almost completed, identifying at least one target gene of almost all proteins of the three families. Especially, it is notable that we found that YsiA protein was involved in global fatty acid degradation regulation. These results of the five-year project, unveiling a broad outline of the transcriptional network of B. subtilis, was published in 39 scientific papers listed below together with more than 100 of presentations at scientific meetings.