Translational control of ROS Management

ROS 管理的翻译控制

• 批准号: 9063543
• 负责人: Thomas J Begley
• 金额: $ 38.73万
• 依托单位: SUNY POLYTECHNIC INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-15 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9063543
• 关键词: Address; Affect; Anticodon; Antioxidants; Automobile Driving; Biochemical; Cancer Model; Cell Death; Cell Proliferation; Cell model; Cell physiology; Cells; Codon Nucleotides; Colon Carcinoma; Colorectal Cancer; Computer Analysis; DNA; DNA Damage; DNA Repair; DNA Sequence Alteration; Data; Down-Regulation; Drug Metabolic Detoxication; Engineering; Enzymes; Equilibrium; Event; Gene Expression; Genes; Genome Stability; Goals; Health; Human; Hydrogen Peroxide; Link; Lipids; Malignant Neoplasms; Malignant neoplasm of urinary bladder; Mass Spectrum Analysis; Measures; Messenger RNA; Methylation; Methyltransferase; Modeling; Modification; Molecular; Monitor; Mutation; Normal Cell; Output; Oxidative Stress; Pattern; Positioning Attribute; Process; Proteins; Publishing; RNA; Reactive Oxygen Species; Reading; Resistance; Resources; Ribonucleotide Reductase; Selenocysteine; Signal Transduction; Signaling Protein; Solid Neoplasm; Stress; Substrate Specificity; System; Terminator Codon; Testing; Transfer RNA; Translations; Tumorigenicity; Work; biological adaptation to stress; cancer cell; cancer prevention; cancer therapy; carcinogenesis; carcinogenicity; disease-causing mutation; glutathione peroxidase; improved; insight; novel; overexpression; prevent; programs; response; thioredoxin reductase; tool; treatment strategy; tumor growth

DESCRIPTION (provided by applicant): Both environmental and endogenous processes generate pathologically high levels of ROS, with subsequent damage to lipids, proteins and DNA promoting mutation and cell death. However, low levels of ROS are physiologically important in regulating cell proliferation and gene expression.1,2 Cells must thus prevent the detrimental and promote the beneficial aspects of ROS, which suggests that there are precise regulatory strategies to maintain proper ROS levels. We propose to test the hypothesis that cells manage ROS balance by turning-on and turning-off the translation of ROS detoxification and DNA damage response proteins, using the opposing activities of the RNA methyltransferases ALKBH8 and TRM9L. The underlying mechanism arises from our discovery that enzyme-catalyzed methylation signals by ALKBH8, operating at the level of tRNA modification to anticodons, can translationally up-regulate selenocysteine (Sec)-containing ROS management proteins. These RNA modifications are required for "stop-codon recoding" - the mechanism of reading internal stop-codons as Sec-codons in mRNAs for Sec-containing glutathione peroxidases (GPXs, antioxidants) and thioredoxin reductases (TrxRs, regulate ribonucleotide reductase activity to promote the DNA damage response). We hypothesize that dueling RNA modifications control stop-codon recoding to promote (ALKBH8) or prevent (TRM9L) the translation of Sec-proteins and thus regulate ROS and DNA damage responses. Further, we hypothesize that part of the process of carcinogenicity involves hijacking of RNA modification systems to turn-off stress responses and promote a pro-mutagenic program of ROS-induced DNA damage and decreased DNA repair. To facilitate our studies, we have developed a modification analysis platform to monitor 33 human tRNA modifications and we have demonstrated that anticodon modifications are significantly regulated in response to ROS stress. Together with our RNA modification analysis platform, we will use biochemical, molecular and computational analyses to test our hypothesis in a robust set of normal, cancer- derived and engineerable lines. Importantly, our studies will define a novel system of translational control of stress response, dissect the mechanism a pro-mutagenic program, and reveal new targets and strategies for cancer treatment and prevention.

描述（由申请人提供）：环境和内源性过程都会产生病理高水平的ROS，随后对脂质，蛋白质和DNA损害，促进突变和细胞死亡。然而，低水平的ROS在调节细胞增殖和基因表达上在生理上很重要。1,2细胞必须防止有害并促进ROS的有益方面，这表明存在确切的调节策略以维持适当的ROS水平。我们建议通过使用RNA甲基转移酶ALKBH8和TRM9L的相对活性来测试细胞通过转动和关闭DNA损伤反应蛋白的翻译来控制ROS平衡的假设。基本机制是我们发现的，酶催化的AlkBH8催化甲基化信号以tRNA修饰水平为反登起，可以翻译上调节含有ROS Management蛋白的硒常生态半胱氨酸（SEC）。这些RNA修饰是“停止二氧化碳的重新编码”所必需的 - 将内部止动物读取为mRNA中的SEC-codons的机制，用于含SEC的谷胱甘肽过氧化物酶（GPXS，抗氧化剂）和硫氧还蛋白还原酶（TRXRS）（TRXRS，调节核糖核苷酸还原酶活性以促进DNA损害响应）。我们假设Dueling RNA修饰控制止动型孔子，以促进（ALKBH8）或预防（TRM9L）SEC蛋白的翻译，从而调节ROS和DNA损伤反应。此外，我们假设致癌过程的一部分涉及劫持RNA修饰系统以关闭应力反应，并促进促进ROS诱导的DNA损伤和DNA修复减少的促症状程序。为了促进我们的研究，我们开发了一个修改分析平台来监测33个人类tRNA修饰，我们证明了反应响应ROS应激的反应密切调节。与我们的RNA修改分析平台一起，我们将使用生化，分子和计算分析来在一组强大的正常，癌症衍生和可工程的线路中测试我们的假设。重要的是，我们的研究将定义一个新型的压力反应控制，剖析机制促症状程序，并揭示癌症治疗和预防的新目标和策略。