Investigation of Comprehensive Vision of Transcriptional regulation in Bacillus subtilis

我们获得了500多个枯草芽孢杆菌DNA微阵列数据，利用不同培养条件和多种环境胁迫下生长的细胞，对改变基因进行基因聚类，并利用转录调控突变体，提取转录调控因子的靶候选基因。通过基因聚类和目标候选基因的鉴定，揭示了一个与支链氨基酸生物合成有关的代谢调控基础网络，包括CcpA、CodY和TnrA对碳氮的调控，以及RelA的严格控制。基于DNA结合转录调控因子的DNA微阵列数据，我们完成了ECF-sigma因子的全面分子遗传学分析，并基本完成了34个双组分调控系统的分子遗传学分析。因此，只有几个双组分系统的功能仍然是未知的。此外，我们在200多个蛋白质中对HTH-DNA结合调节蛋白的MarR、TetR和MerR家族进行了分子遗传学分析，对这些蛋白质的DNA微阵列分析基本完成，在这三个家族中几乎所有蛋白质都至少鉴定出一个靶基因。尤其值得注意的是，我们发现YsiA蛋白参与了脂肪酸降解的全局调控。这个为期五年的项目的这些结果揭示了枯草芽孢杆菌转录网络的大致轮廓，发表在下面列出的39篇科学论文中，以及100多篇科学会议上的报告。

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.