STRUCTURAL ROBUSTNESS OF THE RIBOSOME

核糖体的结构稳健性

• 批准号: 8361667
• 负责人: Gerwald Jogl
• 金额: $ 1.1万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361667
• 关键词: Active Sites; Address; Amines; Amino Acid Substitution; Antibiotic Resistance; Antibiotics; Biological; Collaborations; Data Set; Dependence; Evolution; Funding; Grant; Mutation; National Center for Research Resources; Principal Investigator; Process; Protein Biosynthesis; Research; Research Infrastructure; Resolution; Resources; Ribosomes; Site; Source; Streptomycin; Thermus thermophilus; United States National Institutes of Health; Universities; X-Ray Crystallography; base; cost; fitness; resistance mutation; structural biology; thermophilic bacteria

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The ribosome is the universal site of protein synthesis, containing some of the most highly conserved of all biological sequences. Nevertheless, the ribosome is robust to mutation, capable of functioning when chal-lenged with base or amino acid substitutions in its highly conserved functional centers. As major targets of antibiotics, these functional centers are the sites of numerous antibiotic-resistance mutations. While it has been well established that antibiotic-resistance mutations carry a substantial fitness cost, the structural basis for this burden is only now within the scope of our technical ability to investigate. Here, we will use X-ray crystallography of ribosomes from the thermophilic bacterium Thermus thermophilus to address the structural robustness of ribosome active sites and its relationship to biological fitness. In collaboration with Albert Dahl-berg and Steven Gregory at Brown University, we will investigate streptomycin-resistance mutations to ex-amine the mutational robustness of a conserved ribosome functional center that participates in global con-formational changes of the 30S subunit. In collaboration with Frank Murphy, we have already collected dif-fraction data sets for 30S subunit crystals carrying streptomycin-resistance mutations and the streptomycin-dependence mutation G524U. Overall, we have crystals available for 30S subunits carrying several antibi-otic-resistance mutations and we are in the process of preparing a large number of additional ribosome sam-ples. In addition to providing a more complete mechanistic understanding of antibiotic resistance at an un-precedented level of resolution, these efforts are directed towards establishing fundamental principles of ri-bosome structural organization and evolution.

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 核糖体是蛋白质合成的通用部位，包含所有生物序列中最保守的一些。然而，核糖体对突变是强大的，当与其高度保守的功能中心的碱基或氨基酸替代时，核糖体能够发挥功能。作为抗生素的主要靶点，这些功能中心是许多抗生素耐药突变的地点。虽然众所周知，抗生素耐药性突变会带来巨大的适应成本，但这种负担的结构基础现在才在我们的技术能力调查范围内。在这里，我们将使用来自嗜热细菌Thermus thermophilus的核糖体的X射线结晶学来研究核糖体活性部位的结构稳定性及其与生物适合性的关系。我们将与布朗大学的阿尔伯特·达尔-伯格和史蒂文·格雷戈里合作，研究链霉素耐药突变，以验证参与30S亚基全球构象变化的保守核糖体功能中心的突变稳健性。与Frank Murphy合作，我们已经收集了携带链霉素耐药突变的30S亚单位晶体和链霉素依赖突变G524U的不同组分数据集。总体而言，我们已经有了携带几个抗药性突变的30S亚基的晶体，我们正在准备大量额外的核糖体样本。除了在前所未有的解决水平上提供对抗生素耐药性的更完整的机械性理解之外，这些努力旨在建立核糖体结构组织和进化的基本原则。