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. !

项目总结

项目成果

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

血浆蛋白酶裂解并储存在血小板α颗粒中：单核细胞激活中的潜在作用。

• DOI: 10.1002/rth2.12429
• 发表时间: 2020-10
• 期刊: Research and practice in thrombosis and haemostasis
• 影响因子: 4.6
• 作者: Won E;Morodomi Y;Kanaji S;Shapiro R;Vo MN;Orje JN;Thornburg CD;Yang XL;Ruggeri ZM;Schimmel P;Kanaji T
• 通讯作者: Kanaji T

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

• DOI: 10.1073/pnas.2012898118
• 发表时间: 2021-03-30
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Sun L;Wei N;Kuhle B;Blocquel D;Novick S;Matuszek Z;Zhou H;He W;Zhang J;Weber T;Horvath R;Latour P;Pan T;Schimmel P;Griffin PR;Yang XL
• 通讯作者: Yang XL

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

• DOI: 10.1038/s41423-019-0331-0
• 发表时间: 2021-06
• 期刊: Cellular & molecular immunology
• 影响因子: 24.1
• 作者: Adams RA;Fernandes-Cerqueira C;Notarnicola A;Mertsching E;Xu Z;Lo WS;Ogilvie K;Chiang KP;Ampudia J;Rosengren S;Cubitt A;King DJ;Mendlein JD;Yang XL;Nangle LA;Lundberg IE;Jakobsson PJ;Schimmel P
• 通讯作者: Schimmel P

The endless frontier of tRNA synthetases.

• DOI: 10.1016/bs.enz.2020.09.001
• 发表时间: 2020
• 期刊: The Enzymes
• 作者: Schimmel P