Structural Robustness of Ribosome Functional Centers

 描述(申请人提供)：项目摘要抗药性微生物再次成为世界范围内对人类健康的主要威胁。曾经在对抗致命病原体传播方面至关重要的药物已经变得无效。开发新的抗菌剂来解决这一危机将需要详细了解抗生素耐药性的潜在机制，包括抗生素靶标突变造成的耐药性的结构基础。最重要的抗生素靶标是核糖体，它是蛋白质合成的通用部位，在许多物种中都发现了耐药突变。通过强有力的合作努力，这项提案利用了嗜热热杆菌核糖体的遗传学和X射线结晶学的现状，以解决抗生素耐药性的基本方面。这项建议的目的是：(1)确定在连续暴露于多个抗生素挑战后可能出现的多个抗生素耐药突变之间表型相互作用的结构基础；(2)监测次级突变的进化，以补偿有害的抗生素耐药突变的适应成本。这些目的旨在提供对抗生素耐药性机制的基本见解，研究结果将适用于针对抗生素耐药性病原体的合理药物设计。实现这些目标将涉及遗传学和X射线结晶学的创新融合，以确定突变核糖体的结构。嗜热杆菌特别适合作为模型系统，因为它对基因操作的适应性和它的核糖体适合结晶。我们的建议建立在以下坚实的基础上：(1)在遗传学和X射线结晶学方面拥有丰富的经验，包括实现拟议目标所需技能的经过验证的跟踪记录；(2)广泛的初步结果，包括一些抗生素耐药核糖体的结构确定；(3)从长期的出版物记录中可以看出，在蛋白质合成领域和抗生素耐药性方面具有广泛的工作知识；(4)两家私人投资机构之间有着良好的合作关系。该提案目标的实现将为了解抗生素耐药性的机制提供基本的见解，这将对尚未确定的耐药性突变具有预测能力。它们还将创建一个有价值的框架，用于 新型抗菌剂。

 DESCRIPTION (provided by applicant): Project Summary Antibiotic-resistant microbes are once again a major threat to human health worldwide. Drugs once crucial in fighting the spread of deadly pathogens have become ineffective. The development of novel antimicrobials to solve this crisis will require detailed knowledge of the underlying mechanisms of antibiotic resistance, including the structural basis of resistance caused by mutations in antibiotic targets. The most important antibiotic target is the ribosome, the universal site of protein synthesis, and resistanc mutations have been identified in numerous species. Through a strong collaborative effort, this proposal capitalizes on the current state of genetics and X-ray crystallography of the ribosome from Thermus thermophilus to address fundamental aspects of antibiotic resistance. The aims of this proposal are: (1) determine the structural basis for phenotypic interaction among multiple antibiotic-resistance mutations that can potentially arise after sequential exposure to multiple antibiotic challenges; (2) monitor the evolution of secondary mutations that compensate for the fitness cost of deleterious antibiotic-resistance mutations. These aims are designed to give fundamental insights into mechanisms of antibiotic resistance, and findings will be applicable to the rational design of drugs directed against antibiotic-resistant pathogens. Achieving these aims will involve an innovative fusion of genetics and X-ray crystallography to determine the structures of mutant ribosomes. The bacterium T. thermophilus is especially suited as a model system due to its amenability to genetic manipulation and the suitability of its ribosomes for crystallization. Our proposal rests on a strong foundation of: (1) extensive experience in both genetics and X-ray crystallography, including a proven track record of the skills needed to achieve the proposed aims; (2) extensive preliminary results, including the structure determination of a number of antibiotic-resistant ribosomes; (3) an extensive working knowledge of the protein synthesis field and antibiotic-resistance, as indicated by a long track record of publications; (4) an excellent collaborative relationship between the two PIs. The accomplishment of the goals of this proposal will provide fundamental insights into the mechanism of antibiotic resistance, which will have predictive power regarding resistance mutations not yet identified. They will also create a valuable framework for the rational design of novel antimicrobials.