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球主要的健康问题，目前的疾病预防或治疗策略不足。I型干扰素是一种细胞因子，其通过转录上调数百个基因来诱导细胞中的抗病毒状态。这些基因中的一些编码具有直接抗病毒活性的蛋白质，尽管这些蛋白质中只有一小部分已经被机械地表征。在这些蛋白质中，干扰素诱导的跨膜蛋白3（IFITM 3）最近已被我们和其他人显示对测试的流感病毒的所有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的进入或融合而不是抑制病毒复制来起作用。使用化学报告，我们已经表明，IFITM 3是post-palmitoylated和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IFITM 3也是泛素化的。拟议研究的总体目标是通过表征IFITM 3的作用机制及其通过翻译后修饰的细胞调节来增加对先天抗病毒免疫的理解。在K99阶段，我们将开发生物化学和显微镜检测，以测试棕榈酰化控制IFITM 3的适当运输，使其与病毒颗粒相互作用并防止病毒进入的假设。有了这些知识，我们将在R 00阶段寻找鉴定负责IFITM 3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，我们将应用在K99阶段开发的测定来了解泛素化在IFITM 3生物学中的作用及其与棕榈酰化的相互作用。分析IFITM 3的作用机制和通过一组独特的翻译后修饰控制这种活性可能为利用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）：干扰素诱导型跨膜蛋白3（IFITM 3）已被证明对许多病毒病原体具有广泛的抑制活性。由于目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IFITM 3的运输和与病毒颗粒的相互作用来了解IFITM 3的抗病毒作用机制。