Structural Robustness of Ribosome Functional Centers

核糖体功能中心的结构稳健性

• 批准号: 9266791
• 负责人: Steven Gregory
• 金额: $ 34.73万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-15 至 2020-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9266791
• 关键词: Address; Antibiotic Resistance; Antibiotics; Bacteria; Binding Sites; Biological Models; Complex; Cryoelectron Microscopy; Crystallization; Crystallography; Defect; Development; Distant; Drug Design; Evolution; Exposure to; Financial compensation; Foundations; Funding; Future; Gene Components; Gene Expression; Gene Proteins; Gene Structure; Genes; Genetic; Genetic Translation; Goals; Health; Human; Knowledge; Laboratories; Medical; Microbe; Molecular; Monitor; Mutation; Nature; Organism; Pathogenicity; Pharmaceutical Preparations; Phenotype; Positioning Attribute; Protein Biosynthesis; Proteins; Publications; Publishing; Resistance; Rest; Ribosomal Proteins; Ribosomes; Site; Structure; System; Techniques; Thermus thermophilus; X-Ray Crystallography; antimicrobial; antimicrobial drug; base; cost; design; experience; fighting; fitness; genetic approach; genetic information; genetic manipulation; innovation; insight; mutant; novel; pathogen; public health relevance; rRNA Genes; resistance mechanism; resistance mutation; skills; structural biology; success; thermophilic bacteria; tool

 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.

 描述（由适用提供）：项目摘要抗生素耐药的微生物再次成为全世界人类健康的主要威胁。曾经对致命病原体的传播至关重要的毒品已经无效。解决这一危机的新型抗菌剂的发展将需要详细了解抗生素耐药性的潜在机制，包括由抗生素靶标突变引起的抗性的结构基础。最重要的抗生素靶标是核糖体，蛋白质合成的通用部位，并且在许多物种中都鉴定出了抗性突变。通过强有力的协作努力，该提案利用了热嗜热的核糖体的当前遗传学状态和X射线晶体学，以解决抗生素耐药性的基本方面。该提案的目的是：（1）确定多种抗生素抗性突变之间表型相互作用的结构基础，这些突变可能会在依次暴露于多种抗生素挑战之后可能出现； （2）监测次要突变的演变，以补偿有害抗生素抵抗突变的适应性成本。这些目标旨在提供对抗生素耐药性机制的基本见解，发现将适用于针对抗生素耐药病原体的药物的合理设计。实现这些目标将涉及遗传学和X射线晶体学的创新融合，以确定突变核糖体的结构。细菌T. hytrophilus特别适合于模型系统，因为其对遗传操作的不适及其核糖体对结晶的适合性。我们的建议基于：（1）在遗传和X射线晶体学方面的丰富经验，包括实现拟议目标所需的技能的可靠记录； （2）广泛的初步结果，包括许多抗生素抗性核糖体的结构测定； （3）对蛋白质合成领域和抗生素抗性的广泛工作知识，如长期的出版物记录所示； （4）两个PI之间存在出色的协作关系。实现该提案的目标将提供对抗生素抗性机制的基本见解，抗生素耐药性具有尚未确定的抗药性突变的预测能力。他们还将为合理设计创建一个宝贵的框架 新型抗菌素。