Structural Robustness of Ribosome Functional Centers

项目摘要 核糖体是大分子机器，在整个进化过程中都是保守的，也就是 负责蛋白质的合成，因此是基因的基本组成部分 表情。由于核糖体在生物化学中的核心作用，它已成为 超过一半的抗生素，以及由于基因突变导致的抗生素耐药性的演变 核糖体已成为人类健康的主要威胁。了解病毒的分子基础 因此，抗生素耐药性对抗击肺炎的威胁具有重大意义。 抵抗。在破译抗生素耐药性的机制时，对 还可以获得核糖体功能的机制，从而可能产生新的抗菌剂 药剂通过合理的药物设计。通过持续的协作努力，该提案 利用PI结合遗传学和结构生物学的能力来解决 核糖体结构和功能的基本问题并为未来提供框架 新型抗菌剂的开发。本方案的目的是：(1)对30S亚基进行解剖 TRNA通过核糖体移动过程中的构象动力学；(2)确定两个 易位过程中30-50秒旋转的亚基间桥；(3)确定构造基础 用于通过核糖体的信号通路。 实现这些目标将涉及利用三个绩效指标的专门知识，以产生和 用X射线表征功能有效突变体并确定其结构 结晶学或低温电子显微镜。我们建议的优点包括：(1)广泛性 要应用的实验方法方面的专业知识；(2)长期有效的记录 私人投资机构之间的合作；(3)核糖体领域的广泛出版记录 结构和功能；(4)大量初步数据和正在进行的研究，包括 多项研究接近完成。实现这些目标将极大地推动该领域的发展 通过回答有关抗生素耐药机制的长期问题，虽然 同时扩大了我们对核糖体基本潜在机制的理解。

Project Summary The ribosome is the macromolecular machine, conserved throughout evolution, that is responsible for the synthesis of proteins and is therefore a fundamental component of gene expression. Because of its central role in biochemistry, the ribosome has become the target of more than half of all antibiotics, and the evolution of antibiotic resistance due to mutations in the ribosome has become a major threat to human health. Understanding the molecular basis of antibiotic resistance therefore has significant implications for the fight against the threat of resistance. In deciphering the mechanism of antibiotic resistance, fundamental insights into the mechanism of ribosome function can also be acquired, leading to possible novel antimicrobial agents through rational drug design. Through an ongoing collaborative effort, this proposal capitalizes on the ability of the PIs to combine genetics and structural biology to address fundamental questions of ribosome structure and function and provide a framework for future development of novel antimicrobial agents. The Aims of this proposal are: (1) Dissect 30S subunit conformational dynamics during tRNA movement through the ribosome; (2) Define the role of two intersubunit bridges in 30S-50S rotation during translocation; (3) Determine the structural basis for signaling pathways through the ribosome. Achieving these aims will involve exploiting the expertise of the three PIs to generate and characterize functionally impactful mutants and determine their structures by X-ray crystallography or cryo-electron microscopy. The strengths of our proposal include: (1) extensive expertise in the experimental methods to be applied; (2) a long track record of effective collaboration between the PIs; (3) an extensive publication track record in the field of ribosome structure and function; (4) a large volume of preliminary data and ongoing studies, including multiple studies nearing completion. Accomplishing these goals will significantly advance the field by answering long-standing questions about antibiotic resistance mechanisms, while simultaneously expanding our understanding of the basic underlying mechanism of the ribosome.