Translational regulation during cigarette smoking-induced reprogramming of the tRNA epitranscriptome, in vitro and in a mouse smoking model

体外和小鼠吸烟模型中吸烟诱导的 tRNA 表观转录组重编程过程中的翻译调控

• 批准号: 10376779
• 负责人: Thomas J Begley
• 金额: $ 37.8万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT ALBANY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-09 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10376779
• 关键词: 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone; Alkylating Agents; Alkylation; Anticodon; Aromatic Polycyclic Hydrocarbons; Arsenic; Automobile Driving; Bacteria; Benzo(a)pyrene; Cadmium; Cardiovascular Diseases; Cell Survival; Cells; Chemicals; Chronic Obstructive Pulmonary Disease; Codon Nucleotides; Complex; Complex Mixtures; Cytochrome P450; DNA Damage; Data Set; Disease; Dose; Embryo; Environmental Exposure; Enzymes; Etiology; Exposure to; Family; Fibroblasts; Formaldehyde; Future; Gene Expression; Gene Family; Genes; Genetic Models; Goals; Heat shock proteins; Human; In Vitro; Inflammation; Inflammatory; Knockout Mice; Link; Liver; Lung; Malignant neoplasm of lung; Mammalian Cell; Mediating; Messenger RNA; Metabolic; Metals; Modeling; Modification; Mus; Naphthalene; Nose; Organism; Oxidants; Pathology; Pathway interactions; Pharmaceutical Preparations; Pharmacologic Substance; Play; Proteins; Proteome; Proteomics; RNA; Rattus; Reporter Genes; Ribonucleosides; Role; Smoking; Statistical Data Interpretation; Stress; System; Testing; Tissues; Toxicant exposure; Transcript; Transfer RNA; Translating; Translational Regulation; Translations; Xenobiotics; Yeasts; biological adaptation to stress; cell behavior; cigarette smoke; cigarette smoking; defined contribution; environmental tobacco smoke; environmental tobacco smoke exposure; epitranscriptome; epitranscriptomics; exposed human population; insight; mouse genetics; novel; oxidation; programs; response; selenoprotein; stressor; toxicant

1. PROJECT SUMMARY Cigarette smoking causes cardiovascular disease, chronic obstructive pulmonary disease (COPD), and lung cancer. The etiology of diseases caused by exposure to environmental tobacco smoke (ETS) is confounded by the complexity of ETS as a mixture of >4000 chemicals, including direct-acting and metabolically-activated toxicants. While all organisms respond to environmental exposures by regulating gene expression, we know little about the translational mechanisms linking toxicant exposure to cell survival and disease. Here we seek to understand the role of the dozens of modified ribonucleosides in tRNA – the tRNA epitranscriptome – in regulating the cell response to ETS exposure. Using a unique computational and analytical platform, we have shown that (1) toxicant exposures cause signature changes in the epitranscriptome of yeast and mammalian cells, and (2) that toxicant-induced reprogramming of the tRNA epitranscriptome regulates protein levels by promoting the selective translation of codon-biased mRNAs from families of stress-response genes in yeast. Our studies in bacteria, yeast and mammalian cells1-7 also show that deficiencies in key epitranscriptomic writer enzymes sensitize cells to killing by specific toxicants due to corrupted translation of stress response proteins. We hypothesize that exposure to the complex mixture of ETS will reprogram the tRNA epitranscriptome to reflect the predominant chemical stressors in ETS and that the altered RNA modifications will regulate cell behavior by selective translation of codon-biased stress response mRNAs. In support of this, we have observed agent-specific epitranscriptome reprogramming in the liver from rats exposed to drugs and toxicants (e.g., arsenic; NTP DrugMatrix), and alkylation- and oxidation-specific signature tRNA modification changes in yeast and human cells. Our epitranscriptomic writer-deficient (Alkbh8-null) mice showed that tRNA modification systems required for translating oxidant-detoxifying selenoproteins are vital to surviving exposure to naphthalene, a P450-activated, oxidant-generating, polycyclic aromatic hydrocarbon (PAH) in ETS. In three aims ranging from in vitro studies in cultured mouse cells to a mouse ETS exposure model, we test the idea that the epitranscriptome and translational regulation play an important role in the cell response to ETS. !

1。项目摘要 吸烟会引起心血管疾病，慢性阻塞性肺疾病（COPD）和肺 癌症。暴露于环境烟草烟雾（ET）引起的疾病病因（ETS）被混淆 ET作为> 4000种化学物质的混合物的复杂性，包括直接作用和代谢激活 有毒物质。尽管所有生物都通过控制基因表达来应对环境暴露，但我们几乎不知道 关于将毒物暴露与细胞存活和疾病联系起来的翻译机制。我们在这里寻求 了解数十种修饰的色带外侧在tRNA中的作用 - tRNA表演组 - 在 调节细胞对ETS暴露的反应。使用独特的计算和分析平台，我们有 表明（1）毒物暴露会导致酵母和哺乳动物的表面转录组的签名变化 细胞，（2）有毒物质引起的tRNA表演组的重编程可通过调节蛋白质水平 促进酵母中应激反应基因家族的密码子偏向mRNA的选择性翻译。我们的 细菌，酵母和哺乳动物细胞的研究还表明，关键的表征作者缺乏 酶对细胞敏感的细胞对特异性毒物杀死，原因是压力反应蛋白的翻译损坏。 我们假设暴露于ETS的复杂组合将重新编程tRNA表演组 为了反映ETS中主要的化学应激源，改变的RNA修饰将调节 通过选择性翻译密码子偏置应力反应mRNA的细胞行为。为此，我们有 观察到的特定于药物和毒物大鼠的肝脏中特定特异性的表面参考组重编程 （例如，砷; NTP药物），以及烷基化和氧化特异性的特异性特异性tRNA修饰变化 酵母和人类细胞。我们的表演分组组作者缺乏症（ALKBH8-NULL）小鼠表明tRNA修饰 翻译氧化氧化氧化氧化硒蛋白所需的系统对于幸存暴露至关重要 萘，一种在ETS中的P450激活，氧化剂生成的，多环芳烃（PAH）。在三个目标中 从培养的小鼠细胞中的体外研究到小鼠ETS暴露模型，我们测试了以下观点。 表面转录组和翻译调节在细胞对ETS的反应中起重要作用。 呢