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.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 核糖体是蛋白质合成的通用部位，其中包含所有生物序列中一些最高度保守的。然而，核糖体对突变具有鲁棒性，当在其高度保守的功能中心中用碱或氨基酸取代时能够发挥功能。作为抗生素的主要靶标，这些功能中心是众多抗生素耐药突变的部位。虽然已经很好地确定抗生素抗性突变具有巨大的适应性成本，但这种负担的结构性基础仅在我们调查技术能力的范围内。在这里，我们将使用嗜热细菌嗜热细菌的核糖体的X射线晶体学来解决核糖体活性位点的结构鲁棒性及其与生物适应性的关系。与布朗大学的阿尔伯特·达尔·伯格（Albert Dahl-Berg）和史蒂文·格雷戈里（Steven Gregory）合作，我们将研究抗霉素抗性突变突变，​​以前胺为参与30s亚基的全球构形变化的保守核糖体功能中心的突变鲁棒性。与弗兰克·墨菲（Frank Murphy）合作，我们已经收集了携带链霉素抗性突变和链霉素依赖性突变G524U的30s亚基晶体的差异数据集。总体而言，我们有30 s亚基的晶体，这些亚基携带了几种抗体抗性突变，并且我们正在准备大量其他核糖体Sampels。除了在非前所未有的分辨率水平上提供对抗生素抗性的更完整的机械理解外，这些努力还针对建立RI-Bosome结构组织和进化的基本原理。