Elucidating the role of peptidyl-tRNA hydrolase activities in translation regulation

阐明肽基-tRNA 水解酶活性在翻译调控中的作用

• 批准号: 2345351
• 负责人: Nora Vazquez-Laslop
• 金额: $ 120万
• 依托单位: University of Illinois at Chicago
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-03-01 至 2028-02-29
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2345351&HistoricalAwards=false
• 关键词: Elucidating; role; peptidyl; tRNA; hydrolase

Synthesis of proteins is a fundamental activity that sustains the life of all organisms. The process of synthesizing proteins inside living cells normally occurs efficiently and smoothly. However, occasionally, and even more so under stress conditions, the protein synthesis machinery encounters problems and stalls. Fortunately, cells are equipped with quality control systems dedicated to overcoming problems in protein synthesis. The multiple components and mechanisms of action of the quality control systems are far from being fully characterized and understood. In this project, the cellular factors and molecular mechanism of a previously unknown critical protein synthesis quality control system in bacteria will be studied. The resulting new knowledge will inform new strategies for combating bacterial infections. The research activities will be incorporated in educational programs to train students from high school, college, and graduate programs, as well as postdoctoral researchers. The research findings will be communicated to the scientific community through presentations at local, national, and international meetings and by publications in peer-reviewed journals. It has been assumed that the enzyme peptidyl-tRNA hydrolase (Pth) is essential for bacteria because of its ability to recycle transfer RNA (tRNA) molecules from peptide-tRNA complexes that dissociate from the ribosome during abortive cycles of translation. Recently, it has come to light that Pth is capable of cleaving peptidyl-tRNAs on ribosomes that have stopped making proteins with problematic sequences. This newly discovered involvement of Pth in translation quality control reveals that the reasons for the essentiality of this enzyme are yet to be fully understood. In this project, ribosome profiling (Ribo-Seq), an approach that reveals the positions of ribosomes genome-wide, will help identify the gene sites in two model bacteria (Escherichia coli and Bacillus subtilis) where Pth acts as a rescue factor for ribosomes struggling to translate specific sequences. In addition, biochemical and structural approaches will be used to reveal the mechanistic principles underlying Pth-driven ribosome rescue. The project will advance our understanding of strategies for protein synthesis quality control. The findings of the project may guide future research to design novel antibiotics and treatments for diseases associated with defective protein synthesis.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.

蛋白质的合成是维持所有生物生命的基本活动。在活细胞内部合成蛋白质的过程通常会有效，平稳地发生。但是，偶尔，甚至在压力条件下，蛋白质合成机制会遇到问题和失速。幸运的是，细胞配备了用于克服蛋白质合成问题的质量控制系统。质量控制系统的多种组成部分和作用机制远非充分表征和理解。在该项目中，将研究细菌中先前未知的关键蛋白质合成质量控制系统的细胞因子和分子机制。由此产生的新知识将为您打击细菌感染的新策略。研究活动将纳入教育计划中，以培训高中，大学和研究生课程的学生以及博士后研究人员。研究结果将通过在当地，国家和国际会议上的演讲以及同行评审期刊的出版物传达给科学界。 已经假设，肽基-TRNA水解酶（PTH）对于细菌是必不可少的，因为它的能力从肽中流产周期中从核糖体中解离的肽-TRNA复合物中回收转移RNA（TRNA）分子。最近，已经揭示了PTH能够在核糖体上裂解肽基trnas，这些核糖体停止生产有问题的序列。这种新发现的PTH参与翻译质量控制表明，这种酶的重要性尚未完全了解。在这个项目中，核糖体分析（Ribo-Seq）是一种揭示核糖体基因组位置的方法，将有助于鉴定两个模型细菌（Escherichia coli和bacillus bacillis bacillis）中PTH的基因位点，在PTH中充当了努力转化特定序列的核糖体的拯救因子。此外，将使用生化和结构方法来揭示PTH驱动的核糖体救援的机械原理。该项目将促进我们对蛋白质合成质量控制策略的理解。该项目的发现可能指导未来的研究，以设计与有缺陷蛋白质合成相关的疾病的新型抗生素和治疗方法。该奖项反映了NSF的法定任务，并被认为是值得通过基金会的智力优点和更广泛影响的评估评估标准的评估值得支持的。