Translational Fidelity in Eukaryotes

真核生物的翻译保真度

• 批准号: 7849893
• 负责人: Jonathan D Dinman
• 金额: $ 24.87万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7849893
• 关键词: Amino Acids; Anti-Bacterial Agents; Antiviral Agents; Area; Biochemical; Biological; Biological Process; Cells; Clinical; Collaborations; Communication; Communities; Complex; Disease; Drug Delivery Systems; Drug Design; Elements; Eukaryota; Foundations; Goals; HIV-1; Human; Infection; Link; Maintenance; Malignant Neoplasms; Mediating; Messenger RNA; Modeling; Modification; Molecular; Molecular Genetics; Molecular Models; Mus; Mutation; Organism; Pathway interactions; Patients; Persons; Positioning Attribute; Process; Proliferating; Property; Proteins; RNA; Reading Frames; Research; Research Personnel; Ribosomal Frameshifting; Ribosomal Proteins; Ribosomal RNA; Ribosomes; Saccharomyces cerevisiae; Signal Transduction; Societies; Structure; System; T-Lymphocyte; Terminator Codon; Transfer RNA; Translations; Viral; Virus; Work; Yeasts; abstracting; base; blastomere structure; cancer cell; clinical application; cricket paralysis virus; design; developmental disease; fighting; meetings; molecular modeling; mutant; nanomachine; positional cloning; programs; ribosomal protein L2; ribosomal protein L29; stem; tool

ABSTRACT With the availability of atomic scale structures of ribosomes, the next critical task is to link ribosome structure with biological function. At the biological level, we have been exploring how ribosomes recognize termination codons and maintain translational reading frame using translational recoding signals of viral origin. Viral recoding is important for virus propagation, and their mRNA-based recoding signals have proved to be of great utility in elucidating the molecular mechanisms underlying these essential tasks. These studies have been based on the yeast Saccharomyces cerevisiae model eukaryotic organism because it provides researchers with the most advanced, diverse, and robust toolbox available. Further, we have built upon this foundation to develop a robust and synergistic combination of molecular genetic, biochemical, structural, and molecular modeling tools. This has enabled us to show that both the biophysical interactions between ribosomal proteins, rRNAs and tRNAs, and the biochemical properties of ribosome-associated enzymatic activities are important for proper reading frame maintenance and stop codon recognition. On a broader scale, our work is defining the allosteric communication pathways that connect and coordinate different functional centers of the ribosome with one another. The broad goal of this proposal is to further define how ribosome structure influences function. Aim 1 will determine the effects of targeted mutations on yeast ribosome structure and function. Specifically, reverse genetics approaches will be applied to define the functions of specific ribosomal proteins and ribosomal RNAs. These studies include expansion to examine the effects of two ribosome-associated mutations in mammalian systems. The second aim will characterize the interactions between ribosomes the Cricket Paralysis Virus Internal Ribosome Entry Signal (CRPV IRES), and the HIV-1 programmed -1 ribosomal frameshift signal. The proposed collaborations represent logical expansions of our work into new and exciting areas. We also anticipate that during the course of the proposed studies, breakthroughs will continue to be made in the area of ribosome structure, and that new discoveries relevant to ribosomes and disease will be unveiled. The proposed program will position us to quickly capitalize on these, providing a strong foundation for new and unanticipated discovery opportunities. In the end, this work will make significant contributions to the scientific and clinical communities by both deepening our understanding of the relationship between ribosome structure and function, while broadening our view of translational fidelity and disease. PROJECT NARRATIVE Proliferating cells, be they embryonic cells busily creating new persons, T-cells fighting off infection, or cancer cells overwhelming the patient, absolutely require large numbers of highly accurate ribosomes to meet their needs for synthesis of new proteins. Ribosomes, the central component of this process, are complex biological nanomachines composed of many protein and RNA molecules, and the overall goal of the proposed research is to begin to understand how the atomic scale structure of the ribosome ultimately determines its function. A deeper understanding of the relationship between ribosome structure and function will aid the rational design of new classes of drugs designed to target a diverse array of clinical applications including antiviral and antibacterial agents, as well as drugs targeting a diverse array of cancers, developmental disorders, and other critical diseases afflicting society.

抽象的 随着核糖体原子尺度结构的可用性，下一个关键任务是连接核糖体 具有生物功能的结构。在生物学层面，我们一直在探索核糖体如何识别 终止密码子并使用病毒的翻译重新编码信号维持翻译阅读框 起源。病毒重编码对于病毒传播很重要，其基于 mRNA 的重编码信号已 事实证明，这对于阐明这些基本任务背后的分子机制非常有用。 这些研究基于酿酒酵母模型真核生物 因为它为研究人员提供了最先进、最多样化、最强大的工具箱。更远， 我们在此基础上开发了分子遗传学的强大且协同的组合， 生物化学、结构和分子建模工具。这使我们能够证明 核糖体蛋白、rRNA 和 tRNA 之间的生物物理相互作用，以及 核糖体相关酶活性对于正确阅读框的维护和停止非常重要 密码子识别。在更广泛的范围内，我们的工作正在定义变构通讯途径 核糖体的不同功能中心相互连接和协调。广泛的目标是 该提案旨在进一步定义核糖体结构如何影响功能。目标 1 将确定 靶向突变对酵母核糖体结构和功能的影响。具体来说，反向遗传学 方法将用于定义特定核糖体蛋白和核糖体 RNA 的功能。 这些研究包括扩大检查两种核糖体相关突变的影响 哺乳动物系统。第二个目标将描述核糖体之间的相互作用 麻痹病毒内部核糖体进入信号 (CRPV IRES) 和 HIV-1 编程的 -1 核糖体 移码信号。拟议的合作代表了我们的工作逻辑扩展到新的和 令人兴奋的领域。我们还预计，在拟议的研究过程中，将会取得突破 核糖体结构领域不断取得进展，与核糖体相关的新发现和 疾病将被揭开。拟议的计划将使我们能够快速利用这些，提供 为新的和意想不到的发现机会奠定坚实的基础。最终，这项工作将使 通过加深我们对科学和临床界的理解，做出了重大贡献 核糖体结构和功能之间的关系，同时拓宽我们对翻译保真度的看法 和疾病。项目叙述 增殖细胞，无论是忙碌地创造新人的胚胎细胞，还是战斗的T细胞 摆脱感染，或者癌细胞压倒患者，绝对需要大量 高度准确的核糖体，以满足合成新蛋白质的需求。 核糖体是这一过程的核心组成部分，是复杂的生物体 由许多蛋白质和RNA分子组成的纳米机器，总体目标 拟议的研究是要开始了解原子尺度结构如何 核糖体最终决定其功能。对关系有更深入的了解 核糖体结构和功能之间的关系将有助于合理设计新类别的核糖体 旨在针对多种临床应用的药物，包括抗病毒和 抗菌剂以及针对多种癌症的药物， 发育障碍和其他困扰社会的重大疾病。