CAREER: Mechanisms of RNA-mediated control of gene expression in bacteria

职务名称：职业：RNA介导的细菌基因表达调控机制细菌已经进化出多种方式来感知它们的环境并对变化的条件（包括温度）做出反应。一些细菌进化出的感知温度的策略之一是在信使RNA中放置温度响应序列。这些“RNA温度传感器”会根据温度变化改变其结构，从而控制特定蛋白质的产生。该项目将研究温度敏感性的结构要求，并阐明影响细菌毒力的RNA-RNA相互作用。本研究将报道RNA介导机制中普遍存在的重要过程，包括解链和退火。这一工作将为RNA分子识别和RNA结构与功能的关系提供信息。对RNA结构、稳定性和分子间相互作用的进一步了解将有助于新的基于RNA的生物传感器的工程设计。该项目还将整合研究和教育，以促进高中，本科和研究生的科学培训和教育。作为该项目的一部分，本科生将接受尖端生物分子核磁共振（NMR）光谱学的培训，并将为包含RNA化学位移信息的公共数据库做出贡献。旨在吸引来自传统上代表性不足背景的青年学生参与的外联活动将包括演示磁共振成像技术。该项目的首要科学目标是深入了解细菌病原体L。单核细胞增多症独立地和一致地起作用以控制基因表达。RNA热传感器控制关键转录激活因子PrfA的翻译，PrfA最终控制一组毒力基因的表达。目的1将揭示生物化学和生物物理特性，在RNA温度传感器的温度传感。目的2将表征，在原子的细节，在prfA RNA温度传感器在不同温度下发生的结构和构象的变化。目的3将阐明RNA温度传感器和代谢物结合核糖开关之间的结构和分子联系，它们共同介导毒力基因的表达。为了实现这些目标，我们将使用多学科的方法，结合生物化学，生物物理学和结构技术，包括NMR光谱，小角X射线散射和X射线晶体学。该项目将使人们更深入地了解某些致病细菌如何感知和响应温度信号，以及RNA分子如何重塑和相互作用以控制基因表达。该项目由生物科学理事会分子和细胞生物科学部的分子生物物理学和遗传机制集群支持。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Title: CAREER: Mechanisms of RNA-mediated control of gene expression in bacteriaBacteria have evolved multiple means to sense their environment and respond to changing conditions, including temperature. One strategy that some bacteria have evolved to sense temperature is the placement of temperature-responsive sequences in messenger RNAs. These ‘RNA thermosensors’ change their structure in response to changing temperature, exerting control over the production of particular proteins. This project will investigate the structural requirements for temperature sensitivity and elucidate RNA-RNA interactions that impact bacterial virulence. This research will report on ubiquitous and important process in RNA-mediated mechanisms including melting and annealing. This work will inform on RNA molecular recognition and the RNA structure-function relationship. Further insights into RNA structure, stability, and intermolecular interactions will aid in the engineering of new RNA-based biosensors. This project will also integrate research and education to foster scientific training and education of high school, undergraduate, and graduate students. As a component of this project, undergraduate students will be trained in cutting-edge biomolecular nuclear magnetic resonance (NMR) spectroscopy and will contribute to a public database containing RNA chemical shift information. Outreach activities aimed at engaging young students from traditionally underrepresented backgrounds will include demonstration of magnetic resonance imaging techniques. The overarching scientific objective of this project is to gain insight into how non-coding RNA molecules from the bacterial pathogen L. monocytogenes function independently and in concert to control gene expression. An RNA thermosensor controls the translation of a key transcriptional activator, PrfA, which ultimately controls the expression of a cluster of virulence genes. Objective 1 will reveal the biochemical and biophysical properties that govern temperature sensing in RNA thermosensors. Objective 2 will characterize, in atomic detail, the structure and conformational changes that occur in the prfA RNA thermosensor at various temperatures. Objective 3 will elucidate the structural and molecular link between the RNA thermosensor and a metabolite-binding riboswitch, which function together to mediate expression of virulence genes. To accomplish these objectives, we will use a multidisciplinary approach combining biochemical, biophysics, and structural techniques including NMR spectroscopy, small angle X-ray scattering, and X-ray crystallography. This project will lead to a deeper understanding of how certain pathogenic bacteria sense and respond to temperature signals and how RNA molecules remodel and interact to control gene expression. This project is supported by the Molecular Biophysics and Genetic Mechanism clusters of the Molecular and Cellular Biosciences Division in the Directorate for Biological Sciences.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.