Mechanistic analysis of a posttranslationally modified innate antiviral effector

翻译后修饰的先天抗病毒效应器的机制分析

• 批准号: 8165228
• 负责人: Jacob Yount
• 金额: $ 14万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8165228
• 关键词: Affect; Antiviral Agents; Biochemical; Biological Assay; Biology; Cells; Chemicals; Complex; Cysteine; Data; Development; Disease; Enzymes; Epithelial Cells; Family; Genes; Genetic; Goals; Hand; Health; Hemagglutinin; Host Defense; Immunity; Infection; Influenza; Integral Membrane Protein; Interferon Type I; Interferons; Knowledge; Lipids; Lung; Lysine; Maps; Measures; Membrane; Mentors; Methods; Microscopy; Modification; Natural Immunity; Neurodegenerative Disorders; Outcome; Phase; Physiology; Play; Polyubiquitin; Post-Translational Protein Processing; Prevention strategy; Proteins; Recycling; Regulation; Reporter; Research; Role; Signal Transduction; Site; Testing; Therapeutic; Transcriptional Regulation; Ubiquitination; Vaccine Therapy; Viral; Viral Proteins; Virion; Virus; Virus Diseases; base; cell growth regulation; combat; cytokine; influenzavirus; insight; mutant; novel; palmitoylation; particle; pathogen; prevent; protein acyltransferase; research study; trafficking

DESCRIPTION (provided by applicant): Influenza and other emerging viruses represent major health concerns worldwide, and current strategies for prevention or treatment of disease are insufficient. Type I interferon is a cytokine that induces an antiviral state in cells by transcriptionally upregulating hundreds of genes. Some of these genes encode proteins with direct antiviral activity, though only a small number of these proteins have been mechanistically characterized. Among these proteins, the interferon-inducible transmembrane protein 3 (IFITM3) has been recently shown by us and others to have broad antiviral activity against all subtypes of influenza virus tested as well as a number of other virus families. Furthermore, this protein is unique among interferon effectors in that it appears to act by preventing entry or fusion of viruses rather than inhibiting viral replication. Using chemical reporters, we have shown that IFITM3 is post-translationally palmitoylated and this lipid modification regulates its antiviral activity. Furthermore, our preliminary data indicate that IFITM3 is also ubiquitinated. The overall goal of the proposed research is to increase understanding of innate antiviral immunity by characterizing the mechanism of action of IFITM3 and its cellular regulation by post-translational modifications. In the K99 phase, we will develop biochemical and microscopy assays to test the hypothesis that palmitoylation controls proper trafficking of IFITM3 allowing it to interact with viral particles and prevent viral entry. With this knowledge in hand we will then seek in the R00 phase to identify enzymes responsible for IFITM3 palmitoylation and look at the global transcriptional regulation of palmitoylating enzymes during viral infections. Furthermore, we will apply assays developed in the K99 phase to understand the role of ubiquitination in IFITM3 biology and its interplay with palmitoylation. Analyzing the mechanism of action of IFITM3 and the control of this activity by a unique set of post-translational modifications may provide insights necessary for harnessing the power of type I interferon for combating viral pathogens. PUBLIC HEALTH RELEVANCE (provided by applicant): Interferon-inducible transmembrane protein 3 (IFITM3) has been shown to possess broadly inhibitory activity against a number of viral pathogens. As current therapies and vaccines are insufficient for combating viral disease, we seek to understand the mechanism of antiviral action of IFITM3 by analyzing its trafficking and interactions with viral particles.

描述（由申请人提供）：流感和其他新出现的病毒代表了全世界的主要健康问题，目前预防或治疗疾病的策略还不够。 I 型干扰素是一种细胞因子，通过转录上调数百个基因来诱导细胞抗病毒状态。其中一些基因编码具有直接抗病毒活性的蛋白质，尽管只有少数蛋白质得到了机制表征。在这些蛋白质中，干扰素诱导跨膜蛋白 3 (IFITM3) 最近被我们和其他人证明对所有测试的流感病毒亚型以及许多其他病毒家族具有广泛的抗病毒活性。此外，这种蛋白质在干扰素效应物中是独特的，因为它似乎通过阻止病毒进入或融合而不是抑制病毒复制来发挥作用。使用化学报告基因，我们发现 IFITM3 是翻译后棕榈酰化的，这种脂质修饰调节其抗病毒活性。此外，我们的初步数据表明 IFITM3 也被泛素化。拟议研究的总体目标是通过表征 IFITM3 的作用机制及其翻译后修饰的细胞调节来增进对先天抗病毒免疫的了解。在 K99 阶段，我们将开发生化和显微镜检测方法，以测试棕榈酰化控制 IFITM3 的适当运输，使其与病毒颗粒相互作用并阻止病毒进入的假设。掌握了这些知识后，我们将在 R00 阶段寻找负责 IFITM3 棕榈酰化的酶，并研究病毒感染期间棕榈酰化酶的全局转录调控。此外，我们将应用 K99 阶段开发的检测方法来了解泛素化在 IFITM3 生物学中的作用及其与棕榈酰化的相互作用。分析 IFITM3 的作用机制以及通过一组独特的翻译后修饰对该活性的控制可能为利用 I 型干扰素的力量对抗病毒病原体提供必要的见解。 公共卫生相关性（由申请人提供）：干扰素诱导跨膜蛋白 3 (IFITM3) 已被证明对多种病毒病原体具有广泛的抑制活性。由于当前的疗法和疫苗不足以对抗病毒性疾病，我们试图通过分析 IFITM3 的运输以及与病毒颗粒的相互作用来了解 IFITM3 的抗病毒作用机制。