Molecular analysis of accurate ribosomal translocation

准确核糖体易位的分子分析

• 批准号: 8811631
• 负责人: Kurt L Fredrick
• 金额: $ 32.33万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2018-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8811631
• 关键词: Affect; Amino Acyl Transfer RNA; Antibiotic Resistance; Antibiotics; Anticodon; Bacteria; Beryllium; Binding; Biogenesis; Cells; Chemicals; Codon Nucleotides; Complex; Coupled; Data; Defect; Development; Discrimination; EEF1A1 gene; Elongation Factor; Enzymes; Escherichia coli; Exhibits; Genes; Genetic Epistasis; Goals; Guanosine Triphosphate; Guanosine Triphosphate Phosphohydrolases; Hereditary Disease; Hydrolysis; Inherited; Kinetics; Knowledge; Laboratories; Lead; Light; Link; Malignant Neoplasms; Measures; Mediating; Messenger RNA; Modeling; Modern Medicine; Molecular; Molecular Analysis; Monitor; Multi-Drug Resistance; Mutation; Nature; Organism; Peptide Elongation Factor G; Phenotype; Physiological; Platelet Factor 4; Play; Prevention; Process; Protein Biosynthesis; Proteins; Quality Control; RNA; Reporting; Ribosomal RNA; Ribosomes; Role; Rotation; Sedimentation process; Shoulder; Side; Signal Transduction; Site; Solvents; Structure; Sucrose; Testing; Transfer RNA; Translating; Translation Initiation; Translations; Work; antimicrobial drug; base; combat; density; human disease; insight; mutant; novel; paralogous gene; pathogen; public health relevance; research study; stem; translation factor

DESCRIPTION (provided by applicant): PROJECT SUMMARY In all organisms, proteins are synthesized by ribosomes, large RNA-based enzymes that use aminoacyl- tRNA substrates to translate messenger RNA. In recent years, tremendous progress has been made in elucidating the structure of the ribosome in the absence and presence of substrates and various translation factors. Despite this wealth of structural information, several important questions about the molecular mechanisms of translation remain open. Furthermore, there exist highly conserved protein factors known to interact with the ribosome whose roles in the cell remain unclear. The long-term goal of my laboratory is to fill these gaps in knowledge. Aim 1. Over the past several years, we have been studying mutations in the RNA component (16S rRNA) of the small (30S) subunit that promote miscoding. We found that mutation G299A on the solvent side of the 30S subunit acts by disrupting intersubunit bridge B8, which lies 80 angstroms away. Experiments of Aim 1 will determine which other fidelity mutations (ribosomal ambiguity or ram, error-promoting; restrictive or res, error-reducing) act by allosterically altering B8 and whether codon-anticodon interaction is coupled to B8 disruption. This work will shed light on the conformational dynamics of the subunit that govern the decoding process. Aim 2. In all cells, ribosome assembly / maturation is monitored by quality control mechanisms. These mechanisms are complex and remain incompletely understood. Our recent work on the conserved translational GTPase LepA suggests that the factor contributes to translation initiation, either directly or indirectly. We hypothesize that LepA plays a role in ribosome assembly / maturation, and subunits formed in its absence are functional but defective in initiation. Experiments of Aim 2 will investigate this hypothesis, and may elucidate the physiological role of LepA. Ribosomes are a main target of antibiotics, and defects in translation are associated with a growing number of inherited human diseases and cancers. Insight gained by this project may lead to the development of novel antimicrobial drugs and/or treatments for one or more hereditary diseases.

描述（由申请人提供）： 在所有生物体中，蛋白质的项目摘要都是由核糖体合成的，这些核糖体是使用氨基酰基TRNA底物转换Messenger RNA的大型基于RNA的酶。近年来，在不存在和存在底物和各种翻译因子的情况下阐明核糖体的结构方面取得了巨大进步。尽管有大量的结构信息，但有关翻译的分子机制的几个重要问题仍然开放。此外，存在已知与核糖体相互作用的高度保守的蛋白质因子，其在细胞中的作用仍然不清楚。我实验室的长期目标是在知识中填补这些空白。目的1。在过去的几年中，我们一直在研究促进错误编码的小（30s）亚基的RNA成分（16S rRNA）中的突变。我们发现，30S亚基溶剂侧的突变通过破坏了80埃埃斯特罗姆（Angstroms）的溶解桥间桥梁B8的作用。 AIM 1的实验将确定哪些其他保真度突变（核糖体歧义或RAM，促进错误或RES，限制性或RES，降低错误）通过变构改变B8来起作用，以及密码子 - 抗原相互作用是否与B8中断相关。这项工作将阐明控制解码过程的亚基的构象动态。 AIM 2。在所有细胞中，核糖体组装 /成熟通过质量控制机制监测。这些机制很复杂，并且尚不完全理解。我们最近在保守的翻译GTPase LEPA上的工作表明，该因素直接或间接地有助于翻译起始。我们假设LEPA在核糖体组装 /成熟中起着作用，并且在缺乏的情况下形成的亚基是功能性的，但在起始中有缺陷。 AIM 2的实验将研究这一假设，并可能阐明LEPA的生理作用。核糖体是抗生素的主要靶标，翻译中的缺陷与越来越多的遗传性人类疾病和癌症有关。该项目获得的洞察力可能导致一种新型抗菌药物和/或一种或多种遗传性疾病的治疗方法的发展。