Elucidating the Sensing Abilities of Virulence Regulatory Systems

项目总结 细菌病原体对其蛋白质组进行严格控制。这份提案审查了 肠道沙门氏菌伤寒沙门氏菌是如何发挥这种控制作用的。它直接利用 在上一次赠款期间开发了新的方法和概念突破。 首先，我们将使用荧光识别控制全球dna超螺旋的基因。 一种新的检测DNA的高通量方法--DNA超卷曲的评价 在活细胞中超级盘绕。然后我们将研究已识别的基因是如何控制的 这一必不可少的过程。第二，我们将探索大师通过什么机制 毒力调节剂Phop在以下情况下促进开放阅读框架的转录 结合到完全相同的开放阅读框架。这些位置不同寻常的Phop装订 用一种新的算法--转录因子位点识别发现了这些位点 它捕获全基因组范围内的转录因子结合位点，并克服了 SELEX和CHIP-SEQ的限制第三，我们将确定职位- RNA伴侣CSPC和新的Small的转录机制 蛋白UGTS控制着毒力蛋白UGtL的丰度和活性。这个 拟议的研究将提供对细胞生理学和 很可能是广泛适用的，因为所有的细菌都会超级卷曲它们的DNA，激活基因 转录，并对它们的基因进行转录后控制。

PROJECT SUMMARY Bacterial pathogens exert tight control over their proteomes. This proposal examines how Salmonella enterica serovar Typhimurium exerts this control. It makes direct use of novel methods and conceptual breakthroughs developed during the last grant period. First, we will identify genes that govern global DNA supercoiling using Fluorescence Evaluation of DNA Supercoiling, a novel high throughput method that examines DNA supercoiling in living cells. We will then investigate how the identified genes control this essential process. Second, we will explore the mechanism by which the master virulence regulator PhoP promotes transcription of an open reading frame when binding to that very same open reading frame. These unusually located PhoP binding sites were uncovered with Identification of Transcription Factor Sites, a novel algorithm that captures transcription factor binding sites genome wide and overcomes the limitations of SELEX and ChIP-seq. And third, we will determine the post- transcriptional mechanisms by which the RNA chaperone CspC and the novel small protein UgtS control the abundance and activity of the virulence protein UgtL. The proposed research will provide a molecular understanding of cellular physiology and likely be broadly applicable because all bacteria supercoil their DNA, activate gene transcription, and exert post-transcriptional control of their genes.