Translational quality control by trans-editing domains

通过转编辑域控制翻译质量

• 批准号: 10206957
• 负责人: Karin M Musier-Forsyth
• 金额: $ 38.35万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-17 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10206957
• 关键词: Address; Amino Acids; Amino Acyl-tRNA Synthetases; Aminoglycosides; Area; Bacteria; Cell Survival; Cells; Eukaryota; Family; Genetic Code; Goals; In Vitro; Knowledge; Lead; Physiological; Play; Process; Protein Biosynthesis; Protein Synthesis Inhibitors; Proteins; Quality Control; Research; Ribosomes; Role; Shapes; Solid; Structure; Therapeutic; Transfer RNA; Work; deacylation; design; infancy; knock-down; knowledge base; new therapeutic target; novel; pathogen; proline-tRNA; protein function; therapeutically effective

Summary Aminoacyl-tRNA synthetases (ARSs) establish the rules of the genetic code, whereby each amino acid is attached to a cognate tRNA. Errors in this process lead to mistranslation, which can be toxic to cells. Recent studies suggest that the selective forces exerted by cell-specific requirements and environmental conditions potentially shape quality control mechanisms. Approximately half of the ARSs possess a proofreading (or editing) function to hydrolyze mischarged aa-tRNAs and evidence that non-proteinaceous amino acids pose the greatest threat to fidelity is beginning to emerge. Early work in the Musier-Forsyth lab in the field of translational quality control focused on our discovery of Class II prolyl-tRNA synthetase (ProRS) editing. This led to a detailed mechanistic understanding of the novel bacterial ProRS posttransfer editing domain (INS) and the demonstration that the INS domain, when purified on its own outside the context of the ARS, was fully functional in tRNA deacylation. We subsequently discovered that single-domain INS homologs are widespread in Bacteria and in recent years, our focus in this area has turned almost entirely to understanding the function of these INS-like domains in tRNA editing. However, many open questions regarding the physiological function of these putative trans-editing proteins remain. The overarching goal of the research described in this MIRA application is to uncover the specific functions of a growing family of trans-editing proteins known as the INS superfamily. This diverse yet universally conserved family now has a solid and accumulating in vitro structure-function knowledge base, which strongly supports a role in maintaining translational fidelity. Our knowledge of the broader physiological roles of these proteins, especially in eukaryotes, is still in its infancy and is just beginning to reveal wider roles than previously anticipated. This major gap will be addressed in this work. While classical knock- down screens that only define essential versus non-essential genes do not immediately identify editing domains as essential, the strong conservation of these domains implies they play important, and in most cases still undiscovered, roles in cell survival and competitiveness. Proposed studies are designed to address some of the many open questions with regard to both physiological trans-editing functions and potential moonlighting functions of the INS superfamily. These domains are largely unexplored in eukaryotes, including a novel sub- family cluster that is encoded in many unicellular eukaryotic pathogens. The therapeutic potential of trans-editing domains has not been exploited and represents another major gap in the field that we hope to address by our planned studies. In the long term, combining drugs that target novel translational fidelity mechanisms along with known ribosome-targeting protein synthesis inhibitors such as aminoglycosides, may results in more effective therapeutic strategies.

概括 氨基酰基-TRNA合成酶（ARSS）建立了遗传密码的规则，每个氨基酸都是 附着在同源tRNA上。在此过程中的错误导致误导，这可能对细胞有毒。最近的 研究表明，由细胞特异性需求和环境条件施加的选择性力量 潜在地塑造质量控制机制。大约一半的ARS具有校对（或编辑） 水解错误的AA-TRNA的功能，并证明非蛋白酶氨基酸构成最大的氨基酸 对忠诚的威胁开始出现。在翻译质量领域的Musier-Forsyth实验室的早期工作 控制着重于我们发现II类Prolyl-tRNA合成酶（PROR）编辑的控制。这导致了详细的 对新型细菌螺纹的机械理解后编辑结构域（INS）和演示 当ins域在ARS的上下文之外自行纯化时，在tRNA中完全有效 脱酰化。随后，我们发现单域INS同源物在细菌和 近年来，我们在这一领域的重点几乎完全是理解这些Inslik的功能 tRNA编辑中的域。但是，有关这些推定的生理功能的许多开放性问题 跨编辑蛋白仍然存在。此MIRA应用中描述的研究的总体目标是 揭示不断增长的跨编辑蛋白家族的特定功能，称为INS超家族。这 现在，多样化但普遍保守的家庭现在拥有坚实而积累的体外结构功能知识 基地，强烈支持在维持转化保真度中的作用。我们对更广泛的知识 这些蛋白质的生理作用，尤其是在真核生物中，仍处于起步阶段，才刚刚开始揭示 比以前预期的更广泛的角色。这项工作将解决这个主要差距。而古典的敲门 仅定义基本基因与非必需基因的下屏幕无法立即识别编辑域 至关重要的是，对这些领域的强大保护意味着它们很重要，在大多数情况下仍然 未被发现，在细胞生存和竞争力中的作用。拟议的研究旨在解决一些 关于生理跨编辑功能和潜在月光的许多开放问题 INS超家族的功能。这些领域在真核生物中基本上没有探索，包括一个新颖的子 - 在许多单细胞真核病原体中编码的家庭簇。跨编辑的治疗潜力 域尚未被利用，代表了我们希望通过我们的领域的另一个主要差距 计划研究。从长远来看，将靶向新型翻译保真度机制的药物以及 已知的核糖体靶向蛋白合成抑制剂（如氨基糖苷）可能会导致更有效 治疗策略。