Defining the Role of Aminoacyl-tRNA Synthetases in Human Health and Disease

定义氨酰基-tRNA 合成酶在人类健康和疾病中的作用

• 批准号: 10250308
• 负责人: Anthony Antonellis
• 金额: $ 38.24万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10250308
• 关键词: Address; Affect; Alleles; Amino Acids; Amino Acyl-tRNA Synthetases; Area; Biochemical; Biological Assay; Biological Models; Cell Survival; Data; Diagnosis; Disease; Dominant-Negative Mutation; Enzymes; Family; Genes; Health; Hereditary Disease; Human; In Vitro; Mammalian Cell; Mass Spectrum Analysis; Messenger RNA; Molecular; Mus; Mutagenesis; Neuropathy; Onset of illness; Pathogenicity; Patients; Peripheral Nervous System Diseases; Phenotype; Population; Process; Production; Proteins; Role; Sum; Testing; Tissues; Transfer RNA; Transfer RNA Aminoacylation; Translations; Variant; Yeasts; clinical phenotype; disease phenotype; gain of function; gene function; human disease; improved; in vivo; insight; loss of function; molecular pathology; mouse model; patient population; protein expression; ribosome profiling; therapeutically effective

PROJECT ABSTRACT Aminoacyl-tRNA synthetases (ARSs) are a ubiquitously expressed, essential class of enzymes responsible for ligating amino acids to cognate tRNA molecules. Importantly, 34 of the 37 loci encoding an ARS have been implicated in myriad dominant and recessive clinical phenotypes, making these enzymes a major contributor to human inherited disease. It is now important to systematically assess the role of ARS alleles in human disease phenotypes and to determine how they affect protein translation. These data will provide insight into the molecular pathology of disease-associated ARS alleles, which affect a wide range of tissues. Furthermore, defining the molecular mechanisms of ARS-associated disease will allow rapid patient diagnosis through distinguishing pathogenic from non-pathogenic alleles in human populations. We and others have shown that disease-associated ARS alleles cause a loss-of-function effect on tRNA charging. However, a number of critical questions remain, including: What is the full spectrum of disease phenotypes caused by ARS alleles? What is the subset of ARS alleles in human populations that are pathogenic? How do loss-of-function missense ARS variants cause dominant peripheral neuropathy? and How do loss-of-function ARS variants affect mRNA processing and protein expression? Here, we employ multiple established and complementary model systems—computational, biochemical, cellular, yeast, worm, and mouse—to address the above questions. Our efforts will include: (1) studying patient populations to implicate newly identified ARS variants in disease onset; (2) deeply interrogating ARS-related phenotypes using worm and mouse models; (3) systematically determining the effect of ARS variants on gene function using massively parallel mutagenesis and mammalian cell viability assays; (4) testing neuropathy-associated ARS variants for both dominant- negative and toxic gain-of-function effects in vitro and in vivo; and (5) testing loss-of-function, disease- associated ARS variants for an effect on protein translation via ribosomal profiling and mass spectrometry in yeast, worm, and mouse models. In sum, the areas of study outlined in this proposal will dramatically improve our understanding of how certain ARS alleles give rise to dominant and recessive human disease phenotypes.

项目摘要 氨酰基-tRNA 合成酶 (ARS) 是一类普遍表达的重要酶，负责 重要的是，编码 ARS 的 37 个基因座中的 34 个已被连接。 涉及多种显性和隐性临床表型，使这些酶成为 现在系统地评估 ARS 等位基因在人类疾病中的作用非常重要。 表型并确定它们如何影响蛋白质翻译。这些数据将提供对蛋白质翻译的深入了解。 疾病相关 ARS 等位基因的分子病理学，影响广泛的组织。 明确 ARS 相关疾病的分子机制将允许通过以下方式快速诊断患者 我们和其他人已经证明，区分人群中的致病性和非致病性等位基因。 与疾病相关的 ARS 等位基因会导致 tRNA 充电功能丧失。 关键问题仍然存在，包括：ARS 等位基因引起的全部疾病表型是什么？ 人类中哪些 ARS 等位基因子集具有致病性？ 错义 ARS 变异会导致显性周围神经病变吗？以及功能丧失的 ARS 变异是如何引起的？ 影响 mRNA 加工和蛋白质表达？在这里，我们采用了多种已建立且互补的方法。 模型系统——计算、生化、细胞、酵母、蠕虫和小鼠——来解决上述问题 我们的工作将包括：(1) 研究患者群体，以找出新发现的 ARS 变异。 疾病发作；(2) 使用线虫和小鼠模型深入研究 ARS 相关表型； 使用大规模并行诱变系统地确定 ARS 变体对基因功能的影响 和哺乳动物细胞活力测定；（4）测试与神经病相关的 ARS 变异的两种显性变异： 体外和体内的负面和毒性功能获得效应；以及（5）测试功能丧失、疾病- 通过核糖体分析和质谱分析相关的 ARS 变体对蛋白质翻译的影响 总之，该提案中概述的研究领域将得到显着改善。 我们对某些 ARS 等位基因如何引起显性和隐性人类疾病表型的理解。