CAREER: Biochemical and Structural Mechanisms Controlling tRNA-Modifying Metalloenzymes

职业：控制 tRNA 修饰金属酶的生化和结构机制

• 批准号: 2339759
• 负责人: Jeffrey Mugridge
• 金额: $ 82万
• 依托单位: University of Delaware
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-03-01 至 2029-02-28
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2339759&HistoricalAwards=false
• 关键词: CAREER; Biochemical; Structural; Mechanisms; Controlling

With the support of the Chemistry of Life Processes (CLP) program in the Division of Chemistry (CHE) and the Established Program to Stimulate Competitive Research (EPSCoR), Jeffrey Mugridge from the University of Delaware is studying the biochemical and structural mechanisms controlling metalloenzymes that install important chemical modifications on transfer RNA (tRNA) molecules. Cellular tRNA molecules are decorated with a huge diversity of chemical modifications that are essential for correctly tuning tRNA structure, stability, and efficient and accurate protein synthesis. Defects in the enzymes and biochemical pathways that install tRNA modifications are linked to a wide range of human diseases from cancers to neurodegenerative disorders. However, for many of the enzymes that install key chemical modifications on tRNA, a clear understanding of how these enzymes selectively carry out their modification reactions is missing. Filling this gap in knowledge is important for our fundamental understanding of cellular RNA biology. This proposal will define the detailed atomic-level mechanisms for two distinct classes of metal-dependent enzymes (metalloenzymes) that install key modifications on tRNA that directly impact protein synthesis. Closely integrated with this research, a course-based undergraduate research experience (CURE) will be developed that engages both undergraduates and high-school students from underrepresented groups in a collaborative, discovery-based course where students will make and experimentally test predictions about protein-tRNA interactions. The outcomes from this work could provide new information on how tRNA-modifying metalloenzymes carry out complex, multistep reactions on tRNA and broaden engagement of students in research-focused activities at the undergraduate and high-school levels.Hypermodification of the tRNA anticodon loop is essential for proper codon-anticodon recognition and mRNA decoding in the ribosome during translation. The cell installs numerous, chemically complex modifications at these locations to ensure and control translational efficiency and fidelity. This project combines techniques from biochemistry, structural biology, biophysics, and chemical biology to study two different classes of tRNA-modifying metalloenzymes that install sequential modifications on the anticodon loop of tRNA. The goals of this work are to: (1) define the stepwise chemical mechanisms used by these classes of metalloenzymes to install anticodon loop modifications, (2) unveil the structural mechanisms and protein conformations that control tRNA recognition and modification selectivity, and (3) provide new, broad information about how these classes of metalloenzymes are regulated in the cell. The research outcomes could deepen our basic understanding of tRNA biology and pave the way for future therapeutic development targeting these or similar metalloenzymes and tRNA modification pathways.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.

在化学过程的化学过程（CLP）方案（CHE）和刺激竞争性研究的既定计划（EPSCOR）的支持下，特拉华大学的Jeffrey Mugridge研究了在转移RNA（TRNA）上安装重要化学修饰的生物化学和结构机制。细胞tRNA分子装饰有大量的化学修饰，这些化学修饰对于正确调整tRNA结构，稳定性以及有效且准确的蛋白质合成至关重要。安装tRNA修饰的酶和生化途径中的缺陷与从癌症到神经退行性疾病的广泛人类疾病有关。但是，对于许多在tRNA上安装关键化学修饰的酶，对这些酶如何选择性地进行修饰反应的清晰了解。在知识中填补这一空白对于我们对细胞RNA生物学的基本理解很重要。该建议将定义两种不同类型的金属依赖性酶（金属酶）的详细原子级机制，该酶在直接影响蛋白质合成的tRNA上安装了关键修饰。与这项研究紧密相结合的是，将开发基于课程的本科研究经验（CURE），该研究经验（CURE）与来自代表性不足的小组的本科生和高中生都在合作，基于发现的课程中与学生进行，并在其中学生对蛋白质 - 培训相互作用进行实验测试预测。这项工作的结果可以提供有关如何对tRNA修改金属酶进行复杂的，对tRNA进行复杂的多步反应，并扩大学生在本科和高中级别的研究专注活动中的参与。该单元在这些位置安装了许多化学复杂的修饰，以确保和控制翻译效率和保真度。该项目结合了来自生物化学，结构生物学，生物物理学和化学生物学的技术，以研究两种不同类别的tRNA修改金属酶，这些tRNA量化金属酶在tRNA的反密码子环上安装顺序修饰。这项工作的目的是：（1）定义这些类别的金属酶使用的逐步化学机制来安装反对多子环修饰，（2）揭示控制tRNA识别和修饰选择性的结构机制和蛋白质构象，以及（3）提供有关这些类别的金属enzemes的新的，可提供的新的，可调节的细胞。研究结果可以加深我们对TRNA生物学的基本了解，并为针对这些或类似的金属酶和tRNA修饰途径的未来治疗发展铺平道路。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知识分子和更广泛影响的评估来通过评估来获得支持的。