Ribosome Quality Control Mechanisms in Gram-positive Bacteria

革兰氏阳性细菌的核糖体质量控制机制

• 批准号: 10674019
• 负责人: Heather Feaga
• 金额: $ 36.69万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10674019
• 关键词: Address; Antibiotic Resistance; Antibiotics; Area; Bacillus anthracis; Bacillus subtilis; Biochemical; Cell physiology; Cells; Code; Data; Event; Gene Expression; Genetic; Goals; Gram-Negative Bacteria; Gram-Positive Bacteria; Infection; Knowledge; Laboratories; Modeling; Pathway interactions; Physiological; Process; Protein Biosynthesis; Protein Truncation; Proteome; Quality Control; Regulation; Research; Ribosomes; Terminator Codon; Translational Regulation; Translations; United States; biological adaptation to stress; flexibility; interest; premature; prevent; programs

Project Summary: Although bacterial ribosomes have been biochemically interrogated for decades, unknown mechanisms of regulation and quality control are regularly uncovered and targeted with new antibiotics. Major differences in translation regulation between Gram-positive and Gram-negative bacteria have come to light in recent years. Although more than 200,000 infections per year in the United States are caused by antibiotic resistant Gram- positive bacteria, major gaps remain in our understanding of how Gram-positives perform ribosome quality control. To address these gaps, my research program will focus on two major areas. 1) We will identify and characterize strategies used by Gram-positive bacteria to detect and rescue stalled ribosomes and investigate the physiological impacts of ribosome stalling. Preliminary data from my laboratory supports a model in which ribosome stalling in Bacillus subtilis and Bacillus anthracis results in frameshifting and premature translation termination. This process is expected to result in toxic truncated proteins and trigger stress responses. We will investigate this model using genetic, structural, and biochemical approaches. 2) We will determine how ribosome flexibility and atypical translation events can be used by the cell to increase coding capacity. We are particularly interested in how frameshifting and stop codon read-through regulates gene expression and how environmental inputs control this type of regulation. We will also use unbiased high throughput genetics to uncover new mechanisms that prevent stalling and that regulate programmed frameshifting and stop codon read-through.

项目概要： 尽管细菌核糖体已经被生物化学研究了几十年， 监管和质量控制经常被新的抗生素发现和瞄准。重大差异 近年来，革兰氏阳性菌和革兰氏阴性菌之间的翻译调控已被发现。 尽管美国每年有超过20万例感染是由抗生素耐药性革兰氏菌引起的， 虽然革兰氏阳性细菌的核糖体质量与革兰氏阳性细菌的核糖体质量之间存在着很大的差距， 控制为了解决这些差距，我的研究计划将集中在两个主要领域。1)我们将查明和 描述革兰氏阳性菌用于检测和拯救停滞核糖体的策略，并研究 核糖体停滞的生理影响。我实验室的初步数据支持一个模型， 枯草芽孢杆菌和炭疽芽孢杆菌中的核糖体停滞导致移码和过早翻译 终止这一过程预计将导致有毒的截短蛋白质和触发应激反应。我们将 使用遗传、结构和生物化学方法研究该模型。2)我们将确定核糖体 单元可以使用灵活性和非典型翻译事件来增加编码容量。我们特别 感兴趣的是移码和终止密码子通读如何调节基因表达，以及环境如何 输入控制这类调节。我们还将使用无偏见的高通量遗传学来发现新的 防止停滞和调节程序性移码和终止密码子通读的机制。