Stablization of Fragile Human Transfer RNAs

脆弱人类转移 RNA 的稳定

• 批准号: 10199758
• 负责人: PAUL R SCHIMMEL
• 金额: $ 38.7万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-23 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10199758
• 关键词: 3-Dimensional; Alanine-tRNA Ligase; Amino Acids; Amino Acyl-tRNA Synthetases; Aminoacylation; Appearance; Bacteria; Base Pairing; Binding; Binding Proteins; Cells; Defect; Detection; Disease; Elbow; Environment; Enzymes; Evolution; Future; Genes; Genome; Goals; Homeostasis; Human; Laboratories; Link; Metabolism; Mitochondria; Mitochondrial DNA; Mitochondrial Diseases; Mitochondrial Proteins; Molecular Chaperones; Mutation; Myopathy; Neurodegenerative Disorders; Nuclear; Oxidative Phosphorylation; Pathology; Predisposition; Production; Protein Biosynthesis; Proteins; Publishing; RNA Splicing; Recombinant Proteins; Recombinants; Respiration; Stress; Structure; Testing; Therapeutic; Therapeutic Intervention; Transfer RNA; Translations; Variant; Work; base; design; disease-causing mutation; experimental study; insight; mitochondrial genome; mutant; novel; nuclease

Project Summary This proposal is aimed at understanding enough to eventually have a therapeutic intervention for a large group of human mitochondrial diseases associated with transfer RNAs. These diseases arise from mutations in mitochondrial tRNAs that cause instability and susceptibility to nuclease degradation. We focus on stabilizing these mutant tRNAs by identifying and utilizing a pan-specific mitochondrial tRNA binding protein that stabilize the fragile tRNA structure. We have found one such natural protein from the recently discovered ensemble of human tRNA synthetase splice variants. Because defects in mitochondrial (mt) protein synthesis have an immediate impact on cellular metabolism, it is perhaps not surprising that more than 50% of all identified disease-causing mutations in mtDNA are located within mtDNA genes for tRNAs. And yet, these genes constitute only 10% of the mitochondrial genome. A significant number of mt disease-linked mutations, which cause myopathies, neurodegenerative diseases, and multisystemic disorders, are located in the mt tRNAs. Our laboratory has published extensively on aaRSs from bacteria to humans. However, we did not previously have the insight to suggest a path to stabilize disease-causing mutant tRNAs. aaRSs have progressively acquired new domains in evolution. These new domains are dispensable for the aminoacylation function and are mobilized for specific, novel functions outside of translation. We discovered over 250 splice variants (SVs) of human aaRSs. The majority ablate the catalytic activity but retain the novel motifs. Importantly, most are stable as recombinant proteins. Amongst the SVs, we focus on the few that have the appearance of being chaperones for tRNAs, that is, proteins that bind to and stabilize tRNAs, but with a structure- but not sequence-specific recognition of the outside corner (elbow) of the L-shaped tRNA structure. The goal is to select one or more easy- to-purify, stable, recombinant corner- binding splice variants. These will be tested for their ability, when added in trans, to bind defective mutant mitochondrial tRNAs. Another criterion is that the chosen domain can be applied exogenously to cells and enter the mitochondria. In preliminary work we identified at least one SV that fulfills the criteria we established. If successful, this proposal will suggest a new path for therapeutic intervention of the most prevalent human mitochondrial diseases. !

项目概要 该提案旨在充分了解以最终进行治疗干预 用于治疗一大类与转移 RNA 相关的人类线粒体疾病。这些疾病 由线粒体 tRNA 突变引起，导致不稳定和对核酸酶降解的敏感性。 我们专注于通过识别和利用泛特异性线粒体 tRNA 来稳定这些突变 tRNA 稳定脆弱的 tRNA 结构的结合蛋白。我们发现了一种这样的天然蛋白质 最近发现了人类 tRNA 合成酶剪接变体的集合。 因为线粒体 (mt) 蛋白质合成的缺陷会对细胞产生直接影响。 代谢，这也许并不奇怪，超过 50% 的已识别致病突变 mtDNA 中的 tRNA 位于 mtDNA 基因内。然而，这些基因只占基因总数的 10%。 线粒体基因组。大量与 mt 疾病相关的突变会导致肌病， 神经退行性疾病和多系统疾病都位于 mt tRNA 中。我们的 实验室已发表了大量关于从细菌到人类的 aaRS 的文章。然而，我们并没有 先前有洞察力提出了稳定致病突变 tRNA 的途径。 aaRS 在进化中逐渐获得了新的领域。这些新域名是 对于氨酰化功能来说是可有可无的，并且被动员起来用于特定的、新颖的功能 翻译。我们发现了超过 250 个人类 aaRS 的剪接变体 (SV)。大多数消融 催化活性但保留新颖的基序。重要的是，大多数重组蛋白都是稳定的。 在 SV 中，我们重点关注少数看起来是 tRNA 伴侣的 SV， 即，结合并稳定 tRNA 的蛋白质，但具有结构特异性而非序列特异性识别 L 形 tRNA 结构的外角（肘部）。目标是选择一个或多个简单的 纯化、稳定、重组角结合剪接变体。当这些人的能力受到考验时 添加反式，以结合有缺陷的突变线粒体 tRNA。另一个标准是所选择的 结构域可以外源地应用于细胞并进入线粒体。在前期工作中我们 确定了至少一个满足我们制定的标准的 SV。如果成功，该提案将 为最常见的人类线粒体疾病的治疗干预提出了一条新途径。 ！

项目成果

Serum-circulating His-tRNA synthetase inhibits organ-targeted immune responses.

血清循环 His-tRNA 合成酶抑制器官靶向免疫反应。

• 发表时间: 2021
• 影响因子: 24.1
• 作者: Adams, Ryan A;Fernandes;Notarnicola, Antonella;Mertsching, Elisabeth;Xu, Zhiwen;Lo, Wing;Ogilvie, Kathleen;Chiang, Kyle P;Ampudia, Jeanette;Rosengren, Sanna;Cubitt, Andrea;King, David J;Mendlein, John D;Yang, Xiang;N
• 通讯作者: N

Extracellular tyrosyl-tRNA synthetase cleaved by plasma proteinases and stored in platelet α-granules: Potential role in monocyte activation.

胞外酪氨酰-tRNA 合成酶被血浆蛋白酶裂解并储存在血小板 α 颗粒中：在单核细胞活化中的潜在作用。

• 发表时间: 2020-10
• 作者: Won, Eric;Morodomi, Yosuke;Kanaji, Sachiko;Shapiro, Ryan;Vo, My;Orje, Jennifer N;Thornburg, Courtney D;Yang, Xiang;Ruggeri, Zaverio M;Schimmel, Paul;Kanaji, Taisuke
• 通讯作者: Kanaji, Taisuke

CMT2N-causing aminoacylation domain mutants enable Nrp1 interaction with AlaRS.

引起 CMT2N 的氨酰化结构域突变体使 Nrp1 与 AlaRS 相互作用。

• 发表时间: 2021
• 影响因子: 11.1
• 作者: Sun, Litao;Wei, Na;Kuhle, Bernhard;Blocquel, David;Novick, Scott;Matuszek, Zaneta;Zhou, Huihao;He, Weiwei;Zhang, Jingjing;Weber, Thomas;Horvath, Rita;Latour, Philippe;Pan, Tao;Schimmel, Paul;Griffin, Patrick R;Yang, Xiang
• 通讯作者: Yang, Xiang

The endless frontier of tRNA synthetases.

tRNA 合成酶的无尽前沿。

• 发表时间: 2020
• 作者: Schimmel; Paul
• 通讯作者: Paul

Clinical and molecular characterization of novel FARS2 variants causing neonatal mitochondrial disease.

引起新生儿线粒体疾病的新型 FARS2 变异的临床和分子特征。

• 发表时间: 2023-11
• 影响因子: 3.8
• 作者: Chen, Wenqian;Rehsi, Preeya;Thompson, Kyle;Yeo, Mildrid;Stals, Karen;He, Langping;Schimmel, Paul;Chrzanowska;Wakeling, Emma;Taylor, Robert W;Kuhle, Bernhard
• 通讯作者: Kuhle, Bernhard