Structural Robustness of Ribosome Functional Centers

核糖体功能中心的结构稳健性

• 批准号: 10693898
• 负责人: Steven Gregory
• 金额: $ 36.07万
• 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 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10693898
• 关键词: Active Sites; Address; Affect; Amino Acid Substitution; Antibiotic Resistance; Antibiotics; Binding; Binding Sites; Biochemistry; Biological; Capromycin; Codon Nucleotides; Collaborations; Collection; Complex; Cryoelectron Microscopy; Crystal Formation; Data; Dependence; Development; Drug Design; Elements; Equilibrium; Evolution; Future; Gene Expression; Genetic; Genetic study; Goals; Head; Health; Human; Investigation; Laboratories; Methods; Molecular; Molecular Biology; Molecular Conformation; Molecular Machines; Motion; Movement; Mutation; Nature; Outcome; Phenotype; Positioning Attribute; Protein Biosynthesis; Publications; Resistance; Resolution; Ribosomal Proteins; Ribosomal RNA; Ribosomes; Role; Rotation; Signal Pathway; Signal Transduction; Site; Spectinomycin; Streptomycin; Structure; Structure-Activity Relationship; Transfer RNA; Visualization; Work; X-Ray Crystallography; antimicrobial drug; fighting; fitness; insight; mutant; novel; prevent; resistance mechanism; resistance mutation; structural biology

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.

项目摘要 核糖体是大分子机器，在整个进化中保守，即 负责蛋白质的合成，因此是基因的基本成分 表达。由于其在生物化学中的核心作用，核糖体已成为 所有抗生素的一半以上，以及由于突变而引起的抗生素耐药性的演变 核糖体已成为对人类健康的主要威胁。了解分子基础 因此 反抗。在破译抗生素耐药性机制时，对此的基本见解 还可以获取核糖体功能的机制，从而导致新型抗菌素 通过理性药物设计的代理。通过持续的合作努力，该建议 利用PI结合遗传学和结构生物学解决的能力 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。该提案的目的是：（1）剖析30s亚基 通过核糖体运动过程中的构象动力学； （2）定义两个的作用 易位期间30-50s旋转中的亚基间桥梁； （3）确定结构基础 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PI的专业知识来产生和 表征功能影响力突变体并通过X射线确定其结构 晶体学或冷冻电子显微镜。我们提案的优势包括：（1）广泛 实验方法的专业知识； （2）有效的长期记录 PI之间的合作； （3）核糖体领域的广泛出版记录 结构和功能； （4）大量的初步数据和正在进行的研究，包括 多个研究接近完成。实现这些目标将大大推动该领域 通过回答有关抗生素抗性机制的长期问题，而 同时扩展了我们对核糖体基本潜在机制的理解。

项目成果

Structure of Erm-modified 70S ribosome reveals the mechanism of macrolide resistance.

• DOI: 10.1038/s41589-020-00715-0
• 发表时间: 2021-04
• 期刊: Nature chemical biology
• 影响因子: 14.8
• 作者: Svetlov MS;Syroegin EA;Aleksandrova EV;Atkinson GC;Gregory ST;Mankin AS;Polikanov YS
• 通讯作者: Polikanov YS

The C-terminus of ribosomal protein uS4 contributes to small ribosomal subunit biogenesis and the fidelity of translation.

核糖体蛋白 uS4 的 C 末端有助于小核糖体亚基的生物发生和翻译的保真度。

• DOI: 10.1016/j.biochi.2017.05.004
• 发表时间: 2017
• 期刊: Biochimie
• 影响因子: 3.9
• 作者: Kamath,Divya;Allgeyer,BenjaminB;Gregory,StevenT;Bielski,MargaretC;Roelofsz,DavidM;Sabapathypillai,SharonL;Vaid,Nikhil;O'Connor,Michael
• 通讯作者: O'Connor,Michael

Structural basis of Cfr-mediated antimicrobial resistance and mechanisms for its evasion.

Cfr 介导的抗菌素耐药性的结构基础及其逃避机制。

• DOI: 10.1101/2023.09.27.559749
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Aleksandrova,ElenaV;Wu,KelvinJY;Tresco,BenIC;Syroegin,EgorA;Killeavy,ErinE;Balasanyants,SamsonM;Svetlov,MaximS;Gregory,StevenT;Atkinson,GemmaC;Myers,AndrewG;Polikanov,YuryS
• 通讯作者: Polikanov,YuryS

Alterations in the ribosomal protein bL12 of E. coli affecting the initiation, elongation and termination of protein synthesis.

大肠杆菌核糖体蛋白 bL12 的改变影响蛋白质合成的起始、延伸和终止。

• DOI: 10.1016/j.biochi.2020.06.006
• 发表时间: 2020
• 期刊: Biochimie
• 影响因子: 3.9
• 作者: Younkin,AdamD;Gregory,StevenT;O'Connor,Michael
• 通讯作者: O'Connor,Michael

Serial femtosecond X-ray diffraction of 30S ribosomal subunit microcrystals in liquid suspension at ambient temperature using an X-ray free-electron laser.

使用 X 射线自由电子激光对环境温度下液体悬浮液中的 30S 核糖体亚基微晶进行连续飞秒 X 射线衍射。

• DOI: 10.1107/s174430911302099x
• 发表时间: 2013
• 期刊: Acta crystallographica. Section F, Structural biology and crystallization communications
• 作者: Demirci,Hasan;Sierra,RaymondG;Laksmono,Hartawan;Shoeman,RobertL;Botha,Sabine;Barends,ThomasRM;Nass,Karol;Schlichting,Ilme;Doak,RBruce;Gati,Cornelius;Williams,GarthJ;Boutet,Sébastien;Messerschmidt,Marc;Jogl,Gerwald;Dahlber
• 通讯作者: Dahlber