Single-molecule analysis of how birth and death of mRNAs are regulated inside a bacterial cell

单分子分析细菌细胞内 mRNA 的产生和死亡如何调节

• 批准号: 10275999
• 负责人: Sangjin Kim
• 金额: $ 38.7万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10275999
• 关键词: Age; Bacteria; Bacterial Genes; Birth; Cell Nucleus; Cells; Cessation of life; Coupling; DNA-Directed RNA Polymerase; Ecosystem; Environment; Escherichia coli; Foundations; Gene Expression; Gene Expression Regulation; Genetic Transcription; Goals; Health; Heterogeneity; Human; Image; Industrialization; Measures; Messenger RNA; Methods; Modeling; Physiological; Play; Pollution; Process; Production; Proteins; Research; Ribonucleases; Ribosomal Interaction; Ribosomes; Role; Techniques; Translations; Work; bacterial genetics; improved; mRNA Transcript Degradation; microbiome; pathogen; programs; public health relevance; ribonuclease E; single molecule; spatiotemporal

Project Summary Bacteria are everywhere around us, playing critical roles in our health and global ecosystems. Understanding how bacteria thrive is extremely important in keeping our bodies and environments safe and healthy. Bacteria can dynamically adapt to a wide array of conditions by modifying their gene expression program, for example, by boosting the production of proteins necessary for survival and limiting the wasteful production of others. The ultimate goal of my research is to gain an enriched understanding of bacterial gene regulation, thus I study the very fundamental question of how transcription, translation, and mRNA degradation are performed and regulated in physiological settings. Due to the absence of a nucleus in bacterial cells, these three processes occur in the same cytoplasmic volume without clear separations. Therefore, the cellular mechanisms enabling their coordination inside a single cell offer an important foundation for understanding bacterial gene expression programs. Here I describe projects in my group aiming to define the generalizable principles underlying the spatiotemporal coordination of transcription, translation, and mRNA degradation in bacterial cells. We plan to answer the following key questions: (1) What is the mechanism of transcription-translation coupling? (2) How is the interaction between RNA polymerase (RNAP) and ribosome dynamically regulated? (3) How is the rate of mRNA degradation regulated by the age of mRNA and the subcellular localization of RNase E, the major ribonuclease for mRNA degradation in Escherichia coli? We will answer these questions by imaging the dynamics of RNAP, ribosome, and RNase E at the single-molecule level in live cells. Combining these techniques with bacterial genetics, we will identify factors that can modulate the dynamics of RNAP- ribosome interactions and analyze the subcellular heterogeneity in the localization and function of RNase E. Collectively, our work will uncover new mechanistic principles of bacterial gene regulation and generate new methods for measuring, controlling, and modeling gene expression dynamics at the single-molecule level in live cells. The findings from our work have potential applications for a broad range of human health issues, such as promoting healthy microbiomes, killing pathogens, and improving industrial processes to reduce pollution.

项目概要 细菌在我们周围无处不在，在我们的健康和全球生态系统中发挥着关键作用。理解 细菌如何繁殖对于保持我们的身体和环境的安全和健康极其重要。细菌 可以通过修改其基因表达程序来动态适应各种条件，例如， 通过增加生存所需蛋白质的生产并限制其他人的浪费性生产。 我研究的最终目标是加深对细菌基因调控的了解，因此我研究 转录、翻译和 mRNA 降解是如何进行的这一非常基本的问题 在生理环境中受到调节。由于细菌细胞没有细胞核，这三个过程 发生在相同的细胞质体积中，没有明显的分离。因此，细胞机制使 它们在单细胞内的协调为理解细菌基因提供了重要基础 表达程序。在这里，我描述了我的小组中旨在定义可推广原则的项目 细菌转录、翻译和 mRNA 降解的时空协调的基础 细胞。我们计划回答以下关键问题：（1）转录-翻译机制是什么？ 耦合？ （2）RNA聚合酶（RNAP）与核糖体之间的相互作用是如何动态调控的？ (3) mRNA降解速率如何受mRNA年龄和亚细胞定位的调节 RNase E，大肠杆菌中 mRNA 降解的主要核糖核酸酶？我们将通过以下方式回答这些问题 在活细胞的单分子水平上对 RNAP、核糖体和 RNase E 的动态进行成像。组合 通过这些技术与细菌遗传学的结合，我们将确定可以调节 RNAP 动态的因素 核糖体相互作用并分析 RNase E 定位和功能的亚细胞异质性。 总的来说，我们的工作将揭示细菌基因调控的新机制原理并产生新的 在单分子水平上测量、控制和建模基因表达动态的方法 活细胞。我们的工作结果对广泛的人类健康问题具有潜在的应用， 例如促进健康的微生物组、杀死病原体以及改进工业流程以减少 污染。