Interferon-induced IFITM recruitment of ZMPSTE24 blocks viral endocytic entry

干扰素诱导的 IFITM 募集 ZMPSTE24 阻止病毒内吞进入

• 批准号: 9317424
• 负责人: MARTIN E DORF
• 金额: $ 51.79万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-20 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9317424
• 关键词: Address; Affect; Alleles; Animal Model; Antiviral Agents; Arenaviridae; Arenavirus; Binding Proteins; Biological Assay; Biological Models; Cell Nucleus; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Complement; Cytoplasm; DNA Viruses; Data; Dimerization; Early Endosome; Ebola virus; Endosomes; Family member; Generations; Genetic; Histopathology; Host Defense; Human; IFITM1 gene; In Vitro; Infection; Influenza A virus; Insecta; Integral Membrane Protein; Interferon Type I; Interferons; Knock-in Mouse; Lamin Type A; Lysosomes; Mediating; Membrane; Membrane Fusion; Metalloproteases; Modeling; Molecular; Molecular Probes; Mus; Pathway interactions; Patients; Peptide Hydrolases; Phospholipids; Plants; Process; Progeria; Protein Family; Proteins; Proteomics; RNA Viruses; Recruitment Activity; Regulation; Role; Route; Signal Transduction; Site; Specificity; System; Tertiary Protein Structure; Testing; Therapeutic; Vaccinia virus; Variant; Vertebrates; Vesicle; Vesicular stomatitis Indiana virus; Viral; Viral Load result; Virus; Virus Diseases; Virus-like particle; Yeasts; Zinc; cytokine; design; dimer; experimental study; improved; in vivo; insight; late endosome; mortality; novel; overexpression; protein complex; protein expression; protein function; virus envelope

Interferon-induced IFITM recruitment of ZMPSTE24 blocks viral endocytic entry Abstract Viruses must penetrate the cell’s protective phospholipid bilayer to initiate the infectious process. Enveloped viruses enter the cytoplasm by a membrane fusion mechanism often via an endolysosomal route. Interferon- inducible transmembrane proteins (IFITM) were shown to block a broad spectrum of RNA viruses with endocytic entry. However, how IFITM proteins inhibit viral entry is not clear. This application proposes that ZMPSTE24 (abbreviated ZMP), an evolutionarily conserved membrane associated zinc metalloprotease, is a broad-spectrum antiviral effector downstream of IFITM. Preliminary data suggest that type I interferon (IFN) induces IFITM proteins, which in turn recruit and retain ZMP on endosomes. Experiments involving overexpression and genetic deficiency indicate ZMP has antiviral activity against RNA and DNA viruses, including influenza A virus (IAV), vesicular stomatitis virus (VSV) and vaccinia virus (VACV). Direct viral entry assays using virus-like particles suggest ZMP restricts viral endosomal entry. To understand how ZMP functions, this proposal probes the molecular mechanism controlling ZMP cooperation with IFITM proteins. Aim 1 will determine how ZMP and IFITM cooperate and if either can function independently to restrict viral infection. Critical residues involved in ZMP-IFITM interaction and antiviral function are identified. Aim 2 examines the antiviral requirement for ZMP in vivo using an IAV model in zmp-/- mice. Model systems to investigate the role of ZMP deficiency in human cells are proposed. Aim 3 investigates the molecular mechanisms underlying ZMP blockade of viral entry. The role of ZMP in membrane hemifusion will be characterized. Additional studies examine ZMP dimerization and use AP-MS to identify ZMP interacting molecules which regulate antiviral function. In summary, this project will characterize the IFITM-ZMP antiviral signaling cascade and provide an improved understanding of innate antiviral defense.

干扰素诱导的IFITM募集ZMPSTE24阻断病毒内吞进入 摘要 病毒必须穿透细胞的保护性磷脂双层才能启动感染过程。封套的 病毒通过膜融合机制进入细胞质，通常通过内溶酶体途径。干扰素- 可诱导跨膜蛋白(IFITM)被证明能阻断广泛的RNA病毒 内吞进入。然而，IFITM蛋白如何抑制病毒进入尚不清楚。此应用程序建议 ZMPSTE24(简称ZMP)是一种进化上保守的膜相关锌金属蛋白酶。 IFITM下游的广谱抗病毒效应剂。初步数据显示I型干扰素(干扰素) 诱导IFITM蛋白，进而在内体上招募和保留ZMP。涉及到的实验 过表达和遗传缺陷表明ZMP对RNA和DNA病毒具有抗病毒活性， 包括甲型流感病毒(IAV)、水疱性口炎病毒(VSV)和牛痘病毒(VACV)。直接病毒进入 使用病毒样颗粒的分析表明，ZMP限制了病毒内体的进入。要了解ZMP如何 从功能上探讨ZMP与IFITM蛋白协同作用的分子机制。目标 1将决定ZMP和IFITM如何合作，以及两者是否可以独立发挥作用来限制病毒 感染。确定了参与ZMP-IFITM相互作用和抗病毒功能的关键残基。目标2 在ZMP-/-小鼠体内使用IAV模型检测ZMP的抗病毒需求。对系统建模以 研究ZMP缺乏症在人类细胞中的作用。目标3研究分子 ZMP阻断病毒进入的机制。ZMP在膜半灌流中的作用 特色化的。其他研究检查ZMP二聚化并使用AP-MS鉴定ZMP相互作用 调节抗病毒功能的分子。总而言之，本项目将描述IFITM-ZMP抗病毒药物的特征 信号级联并提供了对先天抗病毒防御的更好理解。