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ZMPSTE 24的IFITM募集阻断病毒内吞进入 摘要 病毒必须穿透细胞的保护性磷脂双分子层才能启动感染过程。笼罩 病毒通过膜融合机制进入细胞质，通常经由内溶酶体途径。干扰素- 可诱导的跨膜蛋白（IFITM）被证明可以阻断广谱的RNA病毒， 内吞进入然而，IFITM蛋白如何抑制病毒进入尚不清楚。本申请提出， ZMPSTE 24（缩写为ZMP）是一种进化上保守的膜相关锌金属蛋白酶，是一种具有生物学活性的锌金属蛋白酶。 IFITM下游的广谱抗病毒效应物。初步数据表明，I型干扰素（IFN） 诱导IFITM蛋白，其继而募集ZMP并将其保留在内体上。实验涉及 过量表达和遗传缺陷表明ZMP具有抗RNA和DNA病毒的抗病毒活性， 包括甲型流感病毒（IAV）、水泡性口炎病毒（VSV）和牛痘病毒（VACV）。病毒直接进入 使用病毒样颗粒的测定表明ZMP限制病毒内体进入。了解ZMP如何 功能，该提案探讨控制ZMP与IFITM蛋白质合作的分子机制。目的 1将确定ZMP和IFITM如何合作，以及是否可以独立发挥作用，以限制病毒 感染鉴定了参与ZMP-IFITM相互作用和抗病毒功能的关键残基。目的2 在zmp-/-小鼠中使用IAV模型检查ZMP的体内抗病毒需求。模型系统 研究ZMP缺陷在人类细胞中的作用。目的3研究分子 ZMP阻断病毒进入的潜在机制。ZMP在膜半融合中的作用将是 表征了其他研究检查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。总之，该项目将表征IFITM-ZMP抗病毒药物 信号级联，并提供了先天抗病毒防御的理解。