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: 8525853
• 负责人: Rea M Lardelli
• 金额: $ 5.22万
• 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/8525853
• 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（由申请人提供）：RNA诱导的沉默途径通过诱导翻译抑制和/或激活靶mRNA的降解来调节基因表达。在包括癌症在内的人类疾病中，越来越多地观察到该通路的异常，以致癌 miRNA（称为 oncomir）的发现为代表。该途径需要约 22nt 的小 RNA 分子（称为微 (mi) RNA）来引导 RNA 诱导的沉默复合物 (RISC) 到达目标 mRNA。然而，沉默途径进展所需的许多因素尚不清楚，包括 ATPase MOV10。 MOV10 与含有 miRNA 的复合物相互作用，并被认为是 miRNA 诱导的沉默所必需的，然而，MOV10 的精确分子功能及其 ATP 酶活性仍然未知。此外，MOV10 是 mRNP 重塑因子 Upf1 的密切同源物，Upf1 重塑 RNA-蛋白质复合物，作为 RNA 加工事件（称为无义介导的衰变）的先决条件。我假设 MOV10 在 RNA 诱导的沉默途径中的作用与 Upf1 类似，可以重塑 RNA-蛋白质复合物并促进该途径的进展。本文提出的工作目标是通过以下方式了解 MOV10 在 mRNA 沉默途径中的作用：(1) 确定 RNA 诱导的沉默中需要 MOV10 的步骤；(2) 研究 MOV10 的 ATPase 结构域在内源性 MOV10 靶标中的作用。 MOV10 在人类细胞 mRNA 沉默中的重要性将通过分析 MOV10 耗竭对小 RNA 产生、RISC 成熟、mRNP 翻译抑制和 mRNA 衰减的整体和生化影响来解决。接下来，将通过使用 MOV10 的 ATPase 突变体来测试 mRNP 和 miRNP 的积累来研究 MOV10 的 ATPase 功能，并假设 MOV10 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 沉默中的机械错误如何导致人类疾病提供新的见解。