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: 10597055
• 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/10597055
• 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; Drug Metabolic Detoxication; 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; Link; Liver; Lung; Malignant neoplasm of lung; Mammalian Cell; Maps; Mediating; Messenger RNA; Metabolic; Metals; Modeling; Modification; Mus; Naphthalene; Nose; Organism; Oxidants; Pathology; Pathway interactions; Pharmaceutical Preparations; Pharmacologic Substance; Physical condensation; Play; Proteins; Proteome; Proteomics; RNA; Rattus; Reporter; Ribonucleosides; Role; Smoking; Statistical Data Interpretation; Stress; System; Testing; Therapeutic; 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. 项目概要 吸烟会导致心血管疾病、慢性阻塞性肺病 (COPD) 和肺部疾病 癌症。暴露于环境烟草烟雾 (ETS) 引起的疾病的病因学受到以下因素的影响： ETS 的复杂性是超过 4000 种化学物质的混合物，包括直接作用和代谢激活的化学物质 有毒物质。虽然所有生物体都通过调节基因表达来应对环境暴露，但我们知之甚少 关于有毒物质暴露与细胞存活和疾病之间联系的转化机制。在这里我们寻求 了解 tRNA（tRNA 表观转录组）中数十种修饰核糖核苷的作用 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）有毒物质暴露会导致酵母和哺乳动物表观转录组的特征变化 (2) 毒物诱导的 tRNA 表观转录组重编程通过以下方式调节蛋白质水平 促进酵母中应激反应基因家族的密码子偏倚 mRNA 的选择性翻译。我们的 对细菌、酵母和哺乳动物细胞1-7的研究还表明，关键表观转录组作者的缺陷 由于应激反应蛋白的翻译损坏，酶使细胞对特定毒物的杀伤敏感。 我们假设暴露于 ETS 的复杂混合物中会重新编程 tRNA 表观转录组 反映 ETS 中主要的化学应激源，并且改变的 RNA 修饰将调节 通过选择性翻译偏向密码子的应激反应 mRNA 来改变细胞行为。为了支持这一点，我们有 在暴露于药物和毒物的大鼠肝脏中观察到药物特异性表观转录组重编程 （例如，砷；NTP DrugMatrix），以及烷基化和氧化特异性特征 tRNA 修饰变化 酵母和人体细胞。我们的表观转录组作者缺陷（Alkbh8-null）小鼠表明，tRNA 修饰 翻译氧化解毒硒蛋白所需的系统对于在暴露于环境中的生存至关重要 萘，ETS 中的一种 P450 激活、产生氧化剂的多环芳烃 (PAH)。三个目标 从培养小鼠细胞的体外研究到小鼠 ETS 暴露模型，我们测试了以下观点： 表观转录组和翻译调控在细胞对 ETS 的反应中发挥着重要作用。 ！