The mammalian small RNA machinery and its role in antiviral defenses

哺乳动物小RNA机制及其在抗病毒防御中的作用

• 批准号: 8967559
• 负责人: Benjamin R. tenOever
• 金额: $ 41.83万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-12-01 至 2019-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8967559
• 关键词: Address; Antiviral Agents; Antiviral Response; Area; Arthropods; Biogenesis; Biological Assay; Cell Nucleus; Cells; Cleaved cell; Complex; Cytoplasm; Data; Development; Disease; Embryo; Engineering; Eukaryotic Cell; Fibroblasts; Future; Genome; Grant; Health; Immune response; Infection; Interferons; Mammalian Cell; Mammals; Maps; Masks; Mass Spectrum Analysis; Mediating; Messenger RNA; MicroRNAs; Molecular; Molecular Biology; Mus; Nematoda; Pathogenicity; Pathway interactions; Pattern; Phenotype; Plants; Post-Translational Protein Processing; Poxviridae; Process; Proteins; RNA; RNA Interference; RNA Virus Infections; RNA Viruses; RNA-Induced Silencing Complex; Research; Research Proposals; Role; Signal Transduction Pathway; Small Interfering RNA; Small RNA; Stem cells; System; Therapeutic; Transcript; Transfection; Vaccinia virus; Viral Proteins; Viral load measurement; Virus; Virus Diseases; Virus Replication; antiviral immunity; base; combat; in vivo; novel; pathogen; response; type I interferon receptor; viral RNA

DESCRIPTION (provided by applicant): Understanding the molecular basis for why viruses cause disease is an essential requirement for future development of antiviral therapeutics. While we know a great deal about how viruses are recognized and the various ways in which they are combated, there are still vast areas of cellular antiviral immunity that remain unknown. This research proposal focuses on one such area. The central question being addressed in this proposal is whether small RNAs, or components of the mammalian small RNA machinery, contribute to the cellular response to infection. The importance of this question and the need to address it experimentally can be justified through extensive preliminary data that suggest a novel means to combat virus infection is present in mammalian cells but awaits characterization. In an effort to understand the interplay between the small RNA pathway of mammals and the response to virus infection we have discovered the potency by which the RNA induced silencing complex (RISC) can inhibit virus replication, isolated the first mammalian virus protein which inhibits RISC function, and found that Drosha, a key player in small RNA biogenesis, actively translocates to the cytoplasm during RNA virus infection. Taken together, we believe this research provides compelling evidence for an unappreciated small RNA-mediated antiviral response in mammals. Here we propose to use the virus antagonist to RISC (called VP55) to define the small RNA response to infection in vivo as well as characterize how Drosha participates in this antiviral activity. The overall objective of this proposal is to delineate howthe small RNA pathway contributes to our response to virus infection.

描述（由申请人提供）：了解病毒引起疾病的分子基础是未来开发抗病毒疗法的基本要求。尽管我们对病毒的识别方式以及对抗病毒的各种方式了解很多，但细胞抗病毒免疫仍有大量领域仍不清楚。本研究提案重点关注这样一个领域。该提案要解决的核心问题是小RNA或哺乳动物小RNA机器的组成部分是否有助于细胞对感染的反应。这个问题的重要性以及通过实验解决这个问题的必要性可以通过大量的初步数据来证明，这些数据表明哺乳动物细胞中存在一种对抗病毒感染的新方法，但有待表征。为了了解哺乳动物的小RNA途径与对病毒感染的反应之间的相互作用，我们发现了RNA诱导沉默复合物（RISC）抑制病毒复制的效力，分离了第一个抑制RISC功能的哺乳动物病毒蛋白，并发现小RNA生物发生的关键参与者Drosha在RNA病毒感染期间主动易位到细胞质。总而言之，我们相信这项研究为哺乳动物中未被重视的小 RNA 介导的抗病毒反应提供了令人信服的证据。在这里，我们建议使用 RISC 的病毒拮抗剂（称为 VP55）来定义小 RNA 对体内感染的反应，并描述 Drosha 如何参与这种抗病毒活性。该提案的总体目标是描述小 RNA 途径如何有助于我们对病毒感染的反应。