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: 8601556
• 负责人: Jacob Yount
• 金额: $ 24.9万
• 依托单位: OHIO STATE 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 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8601556
• 关键词: Affect; Antiviral Agents; Biochemical; Biological Assay; Biology; Cells; Cellular Membrane; 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

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.

流感和其他新兴病毒代表着全球的主要健康问题，以及当前的策略 预防或治疗疾病不足。 I型干扰素是一种诱导抗病毒状态的细胞因子 在细胞中通过转录上调数百个基因。这些基因中的一些用直接编码蛋白质 抗病毒活性，尽管这些蛋白质中只有少数是机械特征的。 在这些蛋白中，最近通过 我们和其他人具有广泛的抗病毒活性，以针对所测试的所有流感病毒的亚型以及数量 其他病毒家庭。此外，该蛋白在干扰素效应子中是独一无二的 通过防止病毒的进入或融合，而不是抑制病毒复制。使用化学记者，我们 已经表明，IFITM3在后翻译后是棕榈酰化的，这种脂质修饰调节其抗病毒 活动。此外，我们的初步数据表明IFITM3也是泛素化的。总体目标 拟议的研究是通过表征机制来提高对先天性抗病毒免疫的理解。 通过翻译后修饰的IFITM3及其细胞调节的作用。在K99阶段，我们将 开发生化和显微镜测定法，以检验棕榈酰化控制适当的假设 IFITM3的运输允许其与病毒颗粒相互作用并防止病毒进入。有了这些知识 然后，我们将在R00阶段寻求识别负责IFITM3棕榈酰化的酶并查看 病毒感染期间棕榈酰化酶的全球转录调节。此外，我们会的 应用在K99阶段开发的分析，以了解泛素化在IFITM3生物学及其ITS中的作用 与棕榈酰化的相互作用。分析IFITM3的作用机理和通过 独特的一组翻译后修改可能会提供利用I型力量所需的见解 干扰素打击病毒病原体。