Small RNA Regulation in Bacteria

项目摘要 细菌必须感知并应对快速变化且经常充满压力的环境。他们制定法律 改变细胞结构和功能的基因表达的快速变化是它们生存的关键。在过去二十 多年来，已经非常清楚的是，细菌中基因表达的转录后调控是 普遍存在，代表了一种重要的应激反应策略。小RNA（sRNA）， mRNA和调节转录延伸，翻译和mRNA的稳定性，现在已知是常见的 细菌转录后调节的介质。小RNA（sRNA）的数量从数十到数百， 细菌的基因组和执行不同的调控机制，但其活动的分子细节， 它们在细菌生理学和毒性中的具体作用仍然知之甚少。我的研究小组 在过去的15年里一直致力于解决这些问题。我们的总体方法是将联合收割机 基因组方法与经典遗传学和生物化学研究sRNAs从分子到 在大肠杆菌和沙门氏菌模型系统中的生理水平。我们的工作已经确定了目标调节子 揭示了一些新的sRNA介导的调节的分子机制。我们有 揭示了促进多靶点sRNA调节子分级调节的因素。我们还发现压力 sRNA突变体的反应和代谢表型，将调节的分子机制与 生理结果。在下一个资助期内，我们将建立一个强大而富有成效的基础， 以前的工作，继续研究新的机制，sRNA依赖的调控，在E。杆菌和 沙门氏菌。我们将扩大我们的工作，调查这些调节机制的普遍性，并确定 新的例子一个强大的合作者团队使用一套不同的技术将使我们能够审问sRNA 在全球范围内和在单个RNA分子水平上与靶mRNA的相互作用。这将使我们能够 生成sRNA体内调控的定量模型，并阐明广泛的sRNA调控 网络，以产生比以往任何时候都更精确的sRNA相互作用图。我们设想继续 利用sRNA的研究将分子生物学与细胞生理学联系起来，揭示了对细胞生理学的基本新见解。 微生物生物学

PROJECT SUMMARY Bacteria must sense and respond to rapidly changing and often stressful environments. Their ability to enact rapid changes in gene expression that alter cell structure and function is key to their survival. In the last twenty years, it has become abundantly clear that post-transcriptional regulation of gene expression in bacteria is pervasive and represents an important stress response strategy. Small RNAs (sRNAs) that base pair with mRNAs and regulate transcription elongation, translation, and mRNA stability are now known to be common mediators of bacterial post-transcriptional regulation. Small RNAs (sRNAs) number from tens to hundreds in bacterial genomes and carry out diverse regulatory mechanisms, yet the molecular details of their activities and their specific roles in bacterial physiology and virulence remain poorly understood. My research group has been engaged in research to address these questions for the last 15 years. Our overall approach is to combine genomic approaches with classical genetics and biochemistry to study sRNAs from the molecular to the physiological level in Escherichia coli and Salmonella model systems. Our work has defined the target regulons of several sRNAs, revealing a number of new molecular mechanisms of sRNA-mediated regulation. We have uncovered factors that promote hierarchical regulation of multi-target sRNA regulons. We have also found stress response and metabolic phenotypes for sRNA mutants, linking the molecular mechanisms of regulation to physiological outcomes. In the next funding period, we will build on a strong and productive foundation of previous work to continue investigating novel mechanisms of sRNA-dependent regulation in E. coli and Salmonella. We will extend our work to investigate the prevalence of these regulatory mechanisms and identify new examples. A strong team of collaborators using a diverse set of techniques will allow us to interrogate sRNA interactions with target mRNAs on a global scale and at the level of single RNA molecules. This will allow us to generate quantitative models for in vivo regulation by sRNAs and elucidate an extensive sRNA regulatory network to produce an sRNA interaction map of greater precision than ever before. We envision continuing to use studies of sRNAs to connect molecular biology to cell physiology to reveal fundamental new insight into the biology of microbes.