The Role of TRIM23 in Autophagy Mediated Antiviral Defenses

TRIM23 在自噬介导的抗病毒防御中的作用

• 批准号: 10353335
• 负责人: Michaela Ulrike Gack
• 金额: $ 44.52万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10353335
• 关键词: Amino Acid Motifs; Antiviral Agents; Antiviral Response; Antiviral Therapy; Autophagocytosis; Autophagosome; Binding; Biochemical; Biological; Biological Process; C-terminal; Cell Culture System; Cells; Cytokine Signaling; Data; Defense Mechanisms; Development; Family member; Foundations; Gene Targeting; Guanosine Triphosphate; Guanosine Triphosphate Phosphohydrolases; Herpesvirus 1; Host Defense; Host resistance; Hydrolysis; Immune; Immune signaling; Immunity; In Vitro; Infection; Interferons; Knockout Mice; Knowledge; Laboratories; Link; Lysosomes; Mediating; Microbiology; Molecular; N-terminal; Natural Immunity; Nature; Nerve Degeneration; Organelles; Pathogenesis; Pathway interactions; Phosphorylation; Phosphotransferases; Physiological; Play; Polyubiquitin; Process; Protein Family; Proteins; Regulation; Role; Series; Structure; TBK1 gene; TRIM Motif; Testing; Ubiquitin C; Ubiquitination; Viral; Viral Pathogenesis; Virus; Virus Diseases; Virus Replication; Wild Type Mouse; Work; antimicrobial; antiviral immunity; cell type; cytokine; design; in vivo; insight; mouse model; mutant; pathogen; pathogenic virus; receptor; response; ubiquitin-protein ligase; upstream kinase; viral resistance

PROJECT SUMMARY Autophagy, the process by which cells dispose of cellular contents, has been increasingly appreciated as an important pathway in antiviral host defenses. Intriguingly, recent studies demonstrated that autophagy and the antiviral type I IFN response are intricately connected, and several molecules known to play key roles in IFN-mediated immunity are also important regulators of autophagy. Among these molecules with dual roles in autophagy and IFN-mediated immunity are several TRIM (tripartite motif) protein family members. However, whereas the molecular mechanisms by which TRIM proteins act as antiviral restriction factors or regulate IFN responses have been well characterized, our knowledge about the role of TRIM proteins in autophagy in response to viral infection is still rudimentary. The proposed study builds on a recent discovery by the Gack laboratory that identified TRIM23 as a critical regulator of autophagy-mediated host defense against a broad range of viruses. TRIM23 interacts with and activates the innate immune molecule TBK1, promoting TBK1-mediated phosphorylation of the selective autophagy receptor p62, which ultimately triggers viral clearance and host resistance. Mechanistically, the N-terminal RING E3 ligase of TRIM23 induces atypical non-degradative K27-linked auto-ubiquitination of the C-terminal ARF GTPase domain, which is unique to TRIM23. ARF ubiquitination results in enhanced activity of TRIM23 to hydrolyze GTP, activate TBK1, and mediate virus resistance. TRIM23 depletion or gene-targeting in various cell types showed that TRIM23-mediated autophagy confers antiviral activity against several viruses including HSV-1. Using molecular, biochemical, cell biological and structural approaches combined with virus infection studies, we will define in precise detail how TRIM23 mediates autophagy during viral infection. This study will yield insight into the mechanisms of TRIM23 activation by upstream regulators during viral infection (Aim 1). We will further determine the molecular details of how TRIM23 activates TBK1 and the role the enzymatic activities of TRIM23 – E3 ligase and GTPase – play in TBK1 activation. Finally, we will determine the physiological relevance of TRIM23 for antiviral host resistance and viral pathogenesis using in vitro cell culture systems and infection studies in TRIM23 knockout mice (Aim 2). Our studies will provide a molecular understanding of a key pathway in the autophagy-mediated host defense, which may guide the rational design of new antivirals.

项目摘要 自噬是细胞处置细胞含量的过程，已经是 有趣的是作为抗病毒宿主防御的重要途径。 研究表明，自噬和I型抗病毒药IFN反应很复杂 连接的，几个已知在IFN介导的免疫中起关键作用的分子也是 自噬的重要调节剂。在这些分子中，在自噬中具有双重作用 IFN介导的免疫力是几个修剪（三方基序）蛋白家族成员。然而， 而修剪蛋白作为抗病毒限制因子或 调节IFN响应的表征已经很好，我们对修剪的作用的了解 自噬的蛋白质响应病毒感染仍然是基本的。 拟议的研究以Gack实验室的最新发现为基础 TRIM23是自噬介导的宿主防御的关键调节剂，以针对广泛的范围 病毒。 TRIM23与先天免疫分子TBK1相互作用并激活 选择性自噬接收器p62的TBK1介导的光磷酸化，最终 触发病毒清除和宿主阻力。从机械上讲，N末端环E3连接酶 TRIM23诱导C末端ARF的非典型非降解性K27连接自动泛素化 GTPase域，这是TRIM23所独有的。 ARF泛素化导致 将TRIM23用于水解GTP，激活TBK1并介导病毒抗性。 TRIM23耗竭或 各种细胞类型中的基因靶向表明，TRIM23介导的自噬抗病毒病毒 针对包括HSV-1在内的几种病毒的活性。 使用分子，生化，细胞生物学和结构方法结合 病毒感染研究，我们将精确地定义Trim23如何介导自噬的详细信息 病毒感染。这项研究将洞悉TIRIM23激活的机理 病毒感染期间的上游调节剂（AIM 1）。我们将进一步确定分子 TRIM23如何激活TBK1和TRIM23 - E3的酶促活性的详细信息 连接酶和GTPase - 播放TBK1激活。最后，我们将确定生理 TRIM23对于使用体外细胞的抗病毒宿主耐药性和病毒发病机理的相关性 TRIM23敲除小鼠中的培养系统和感染研究（AIM 2）。我们的研究将提供 对自噬介导的宿主防御中的关键途径的分子理解，该途径 可以指导新抗病毒药的理性设计。