Mechanism and functions of DALRD3-dependent tRNA modification

DALRD3依赖性tRNA修饰的机制和功能

• 批准号: 10706953
• 负责人: Dragony Fu
• 金额: $ 30.8万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10706953
• 关键词: Affect; Alleles; Amino Acyl-tRNA Synthetases; Aminoacylation; Anticodon; Arginine; Binding; Biological; Biology; Brain; Brain Diseases; Cells; Chemicals; Codon Nucleotides; Complex; Development; Developmental Delay Disorders; Disease; Elements; Ensure; Enzymes; Epilepsy; Exhibits; Foundations; Genes; Goals; Health; Human; Individual; Knock-out; Link; Mammalian Cell; Measures; Mediating; Messenger RNA; Methylation; Methyltransferase; Modification; Molecular; Molecular Biology; Molecular Conformation; Monitor; Mouse Strains; Multienzyme Complexes; Mus; Neurodevelopmental Disorder; Neurologic; Organ; Pathogenicity; Pathology; Patients; Physiological; Play; Protein Biosynthesis; Proteins; RNA Biochemistry; RNA Conformation; RNA Folding; RNA Sequences; RNA Stability; RNA methylation; Reporter; Research; Ribosomal Interaction; Ribosomes; Role; Structure; Testing; Therapeutic; Transfer RNA; Translations; Variant; autosome; brain tissue; epileptic encephalopathies; genetic variant; insight; loss of function; nervous system disorder; neurodevelopment; novel; recruit; ribosome profiling

PROJECT SUMMARY Numerous genetic variants in tRNA modification enzymes have been linked to devastating neurodevelopmental and neurological disorders. However, the molecular mechanisms underpinning these pathologies are unknown. Why is it that perturbations to many different tRNA modification enzymes and thus, changes to the various chemical modifications they catalyze, seem to affect the brain more so than other organs? To resolve this question, our lab seeks to elucidate the molecular and cellular roles of tRNA modification enzymes in human health and disease. We have recently uncovered a novel tRNA synthetase-like mimic, DALRD3, that is required for a specific chemical modification in a subset of human tRNAs. Our preliminary results suggest that the DALRD3-dependent modification impacts tRNA conformational stability and function. Notably, we have also identified an autosomal-recessive variant in the DALRD3 gene that causes loss of function and the neurological disorder epileptic encephalopathy. Based upon these findings, we propose that DALRD3-mediated modification plays a critical role in the proper function of specific tRNAs important for protein synthesis during neurodevelopment. In our first Aim, we will define the requirements for tRNA recognition and modification dependent in DALRD3 and its cognate tRNA substrates. For our second Aim, we will measure the impact of DALRD3-dependent modification on tRNA structure and function. In our final Aim, we will determine the role of DALRD3-dependent tRNA modification on global protein translation in the brain through ribosome profiling. In total, the proposed research will have broad implications in understanding how tRNA modifications can impact proper neurodevelopment. Although DALRD3 was an unexpected player in tRNA modification, we now have a new target to explore potential therapeutics for individuals suffering from epileptic encephalopathies linked to tRNA biology.

项目摘要 tRNA修饰酶中的许多遗传变异已与 毁灭性神经发育和神经系统疾病。但是，分子 支撑这些病理的机制尚不清楚。为什么对 许多不同的tRNA修饰酶，因此，各种化学物质变化 他们催化的修改似乎比其他器官更影响大脑？到 解决这个问题，我们的实验室试图阐明tRNA的分子和细胞作用 人类健康和疾病中的修饰酶。我们最近发现了一本小说 tRNA合成酶样模拟物，dalrd3，是特定化学修饰所必需的 在人类trnas的子集中。我们的初步结果表明dalrd3依赖性 修改会影响tRNA构象稳定性和功能。值得注意的是，我们也有 确定了DALRD3基因中的常染色体衰竭变体，导致损失 功能和神经系统疾病癫痫性脑病。基于这些 调查结果，我们建议DALRD3介导的修饰在适当的 特定TRNA对神经发育过程中蛋白质合成很重要的功能。在 我们的第一个目标，我们将定义tRNA识别和修改的要求 依赖于dalrd3及其同源tRNA底物。为了我们的第二个目标，我们将 测量DALRD3依赖性修饰对tRNA结构和功能的影响。 在我们的最终目标中，我们将确定dalrd3依赖性tRNA修饰在 通过核糖体分析，大脑中的全球蛋白质翻译。总共提议 研究将对理解tRNA修改如何可以具有广泛的影响 影响正确的神经发育。虽然dalrd3是tRNA中意外的球员 修改，我们现在有一个新的目标来探索个人的潜在治疗剂 患有与tRNA生物学有关的癫痫性脑病。