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

基本信息

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
项目类别：
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

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

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项目摘要

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.

项目摘要氨基酰基-TRNA合成酶（ARSS）是一种普遍表达的，必不可少的酶类，负责将氨基酸绑扎到同志tRNA分子。重要的是，编码AR的37个基因座中有34个已经以无数占主导地位和隐性临床表型实施，使这些酶成为有助于人遗传疾病。现在，系统地评估ARS等位基因在人类疾病中的作用很重要表型并确定它们如何影响蛋白质翻译。这些数据将为您提供有关疾病相关的ARS等位基因的分子病理，会影响广泛的组织。此外，定义与ARS相关疾病的分子机制将允许通过在人群中区分致病性与非致病等位基因。我们和其他人表明与疾病相关的ARS等位基因对tRNA充电产生功能丧失作用。但是，许多仍然存在关键问题，包括：ARS等位基因引起的全部疾病表型是什么？致病性的人群中ARS等位基因的子集是什么？功能丧失如何错过ARS变体会导致主导的周围神经病？以及功能丧失的ARS变体影响mRNA加工和蛋白质表达？在这里，我们采用了多个建立和完整的模型系统 - 计算，生化，细胞，酵母，蠕虫和鼠标，以解决上述问题。我们的努力将包括：（1）研究患者人群以实施新确定的ARS变体疾病发作；（2）使用蠕虫和小鼠模型深入询问与ARS相关的表型；（3）系统地确定ARS变体对基因功能的影响，使用大量平行诱变和哺乳动物的细胞活力评估；（4）测试与神经病相关的ARS变体，用于两种显性 - 体外和体内的负功能效果负和有毒功能效应；（5）测试功能丧失，疾病 - 相关的ARS变体可通过核糖体分析和质谱法对蛋白质翻译产生影响酵母，蠕虫和鼠标模型。总而言之，该提案中概述的研究领域将显着改善我们对某些ARS等位基因如何产生主导和隐性人类疾病表型的理解。