Structural Robustness of Ribosome Functional Centers

项目摘要 核糖体是大分子机器，在整个进化过程中保持不变， 负责蛋白质的合成，因此是基因的基本组成部分。 表情由于其在生物化学中的核心作用，核糖体已成为 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。了解的分子基础 因此，抗生素耐药性对抗击 阻力在破译抗生素耐药性机制的过程中， 核糖体功能的机制也可以获得，导致可能的新的抗菌剂 通过合理的药物设计。通过持续的合作努力，该提案 利用PI的能力，将联合收割机遗传学和结构生物学结合起来， 核糖体结构和功能的基本问题，并提供了一个框架， 开发新型抗菌剂。本研究的目的是：（1）解剖30 S亚基 tRNA通过核糖体运动过程中的构象动力学;（2）定义两个 易位过程中30 S-50 S旋转中的亚基间桥;（3）确定结构基础 通过核糖体的信号通路。 实现这些目标将涉及利用这三个PI的专业知识， 表征功能上有影响力的突变体，并通过X射线确定其结构 晶体学或低温电子显微镜。我们建议的优点包括：（1）广泛 （2）在实验方法方面的专业知识;（2）长期有效的 PI之间的合作;（3）在核糖体领域的广泛出版记录 结构和功能;（4）大量的初步数据和正在进行的研究，包括 多项研究接近完成。实现这些目标将大大推动该领域的发展 通过回答关于抗生素耐药性机制的长期问题， 同时扩展了我们对核糖体基本机制的理解。

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.