What is the molecular function of MOV10 ATPase in RNA-induced silencing of mRNA?

MOV10 ATPase 在 RNA 诱导的 mRNA 沉默中的分子功能是什么？

• 批准号: 8662561
• 负责人: Rea M Lardelli
• 金额: $ 5.51万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2015-05-17
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8662561
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; ATPase Domain; Address; Binding; Biochemical; Biological Assay; Boxing; Cell physiology; Cells; Complement; Complex; Disease; Event; Gene Expression; Genetic Translation; Goals; Guide RNA; Homologous Gene; Human; Immunofluorescence Immunologic; Lead; Malignant Neoplasms; Mediating; Messenger RNA; MicroRNAs; Molecular; Monitor; Nonsense-Mediated Decay; Oncogenic; Orthologous Gene; Pathway interactions; Play; Process; Production; Proteins; Publications; RNA; RNA Decay; RNA Helicase; RNA Interference; RNA Processing; RNA Stability; RNA-Induced Silencing Complex; Regulation; Reporter; Repression; Role; Small Interfering RNA; Small RNA; Testing; Translational Repression; Translations; Work; base; deep sequencing; helicase; human disease; insight; mRNA Decay; messenger ribonucleoprotein; mutant; novel; novel therapeutics; protein complex; public health relevance; research study; translation assay

DESCRIPTION (provided by applicant): The RNA-induced silencing pathway regulates gene expression by inducing translational repression and/or activating degradation of targeted mRNAs. Aberrations in this pathway are increasingly observed in human diseases, including cancer, as represented by the discovery of oncogenic miRNAs, known as oncomirs. This pathway requires a ~22nt small RNA molecule known as a micro (mi) RNA to guide the RNA-induced silencing complex (RISC) to the targeted mRNA. However, many of the factors that are required for the silencing pathway to progress are not well understood, including the ATPase MOV10. MOV10 interacts with miRNA-containing complexes and is believed to be required for miRNA- induced silencing, however, the precise molecular function of MOV10, and its ATPase activity, remain uncharacterized. Additionally, MOV10 is a close homolog of the mRNP-remodeling factor, Upf1, which remodels RNA-protein complexes as a prerequisite to the RNA-processing event known as nonsense mediated decay. I hypothesize that MOV10 acts analogously to Upf1 during the RNA-induced silencing pathway to remodel the RNA-protein complexes and facilitate progression of this pathway. The goal of the work proposed here is to understand the role of MOV10 in the mRNA silencing pathway by (1) determining the step(s) at which MOV10 is required in RNA-induced silencing and (2) investigating the role of the ATPase domain of MOV10 in the context of endogenous MOV10 targets. The importance of MOV10 in mRNA silencing in human cells will be addressed by analyzing the global and biochemical effects of MOV10 depletion on small RNA production, RISC maturation, translational repression of mRNPs, and mRNA decay. Next, the ATPase function of MOV10 will be studied by using ATPase mutants of MOV10 to test for accumulation of mRNPs and miRNPs with the hypothesis that MOV10 ATPase is required for a remodeling event. Elucidating how MOV10 facilitates RNA-induced silencing will add to our general understanding of mRNA silencing, and is likely to provide novel insights into how mechanistic errors in mRNA silencing may lead to human disease.

描述(申请人提供)：RNA诱导的沉默途径通过诱导翻译抑制和/或激活靶向mRNAs的降解来调节基因表达。这一途径的异常在包括癌症在内的人类疾病中越来越多地被观察到，其代表是致癌miRNAs的发现，即众所周知的oncomir。这一途径需要一个~22nT的小RNA分子，称为微(MI)RNA，以引导RNA诱导的沉默复合体(RISC)与目标mRNA结合。然而，沉默途径取得进展所需的许多因素还不是很清楚，包括ATPase MOV10。MOV10与含有miRNA的复合体相互作用，被认为是miRNA诱导沉默所必需的，然而，MOV10的确切分子功能和它的ATPase活性仍不清楚。此外，MOV10是mRNP重塑因子UPF1的紧密同系物，UPF1重塑RNA-蛋白质复合体，作为RNA处理事件的先决条件，称为无义介导的衰退。我推测，在RNA诱导的沉默途径中，MOV10的作用类似于UPF1，以重塑RNA-蛋白质复合体，促进这一途径的进展。本工作的目的是通过(1)确定MOV10在RNA诱导的沉默中所需的步骤(S)和(2)研究MOV10的ATPase结构域在内源性MOV10靶标中的作用，从而了解MOV10在mRNA沉默途径中的作用。通过分析MOV10缺失对小RNA生产、RISC成熟、mRNPs翻译抑制和mRNA衰退的全局和生化影响，阐述MOV10在人类细胞mRNA沉默中的重要性。接下来，我们将利用MOV10的ATPase突变体来研究MOV10的ATPase功能，以测试mRNPs和miRNPs的积累，假设MOV10的ATPase是重塑事件所必需的。阐明MOV10如何促进RNA诱导的沉默将增加我们对mRNA沉默的一般理解，并可能为mRNA沉默中的机械性错误如何导致人类疾病提供新的见解。