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.

项目摘要 细菌无处不在，在我们的健康和全球生态系统中扮演关键角色。理解 细菌在保持身体和环境的安全和健康方面非常重要。细菌 可以通过修改其基因表达程序，例如，可以动态地适应各种条件 通过增强生存所必需的蛋白质生产并限制他人的浪费。 我研究的最终目的是获得对细菌基因调节的丰富理解，因此我研究 转录，翻译和mRNA降解如何进行的一个非常基本的问题以及 在生理环境中进行调节。由于细菌细胞中没有核，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，启用的细胞机制 它们在单个细胞内的协调为理解细菌基因提供了重要的基础 表达程序。在这里，我在小组中描述了旨在定义可推广原则的项目 细菌中转录，翻译和mRNA降解的时空配位的潜在 细胞。我们计划回答以下关键问题：（1）转录翻译的机制是什么 耦合？ （2）如何动态调节RNA聚合酶（RNAP）和核糖体之间的相互作用？ （3）如何通过mRNA年龄调节mRNA降解速率和 RNase E，大肠杆菌中mRNA降解的主要核糖核酸酶？我们将通过 成像活细胞中单分子水平的RNAP，核糖体和RNase E的动力学。结合 这些具有细菌遗传学的技术，我们将确定可以调节RNAP-动力学的因素 核糖体相互作用并分析RNase E的定位和功能中的亚细胞异质性。 总的来说，我们的工作将发现细菌基因调节的新机械原理，并产生新的机械原理 测量，控制和建模基因表达动力学在单分子水平上的方法 活细胞。我们工作中的发现有潜在的应用于广泛的人类健康问题， 例如促进健康的微生物组，杀死病原体和改善工业过程以减少 污染。