Structural Regulation of IRF3 Activation and Association with Co-activators of the IFN Beta Enhanceosome

IRF3 激活的结构调控及其与 IFN Beta 增强体共激活剂的关联

• 批准号: 10390266
• 负责人: Anne Jecrois
• 金额: $ 0.25万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-09 至 2021-07-16
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10390266
• 关键词: ATF2 gene; Affect; Amino Acids; Antiviral Agents; Antiviral Response; Autoimmune Diseases; Binding; Biochemical; Biological; Biological Assay; Biological Response Modifiers; Cell Nucleus; Cell surface; Cells; Complex; Core Facility; Coupled; Cryoelectron Microscopy; Crystallization; Crystallography; Cytoplasm; DNA; DNA Binding Domain; Dimerization; Emerging Communicable Diseases; Enhancers; Environment; Exposure to; Genes; Genetic Transcription; Homo; Human body; Immune; Immune Response Genes; Immune system; Immunologist; Infection; Inflammation; Interferon Activation; Interferon Suppression; Interferon-beta; Interferons; JUN gene; Length; Light; Macromolecular Complexes; Measures; Mediating; Mentors; Modification; Molecular; Molecular Conformation; Molecular Sieve Chromatography; Molecular Structure; Mutagenesis; Mutation Analysis; Nucleic Acids; Nucleoproteins; Pharmaceutical Preparations; Positioning Attribute; Process; Promoter Regions; Proteins; Regulation; Research; Resolution; Role; Signal Pathway; Signal Transduction; Structure; TNFRSF5 gene; Techniques; Testing; Therapeutic; Training; Transactivation; Transcription Coactivator; Transcriptional Regulation; Variant; Viral; Virus; Virus Diseases; X-Ray Crystallography; base; cytosolic receptor; design; dimer; experimental study; immune activation; insight; light scattering; p65; pathogen; protein complex; protein protein interaction; reconstitution; recruit; response; small molecule; structural biology; transcription factor

PROJECT SUMMARY Following exposure to a virus, the immune system activates and takes rapid measures to protect the body from a spreading infection. Our understanding of the signaling cascade that leads to immuno-activation in response to viral infections is crucial for designing efficient therapeutic drugs that regulate this process. Cell surface and cytosolic receptors recognize pathogen-specific molecules such as viral nucleic acids that activate the interferon regulator factors 3 (IRF3), which is a master regulator of immune activation. Once activated, IRF3 promotes the expression of interferon β (IFNβ), which is essential for the antiviral response. Despite IRF3's active role in antiviral response, dysregulation in IRF3 expression has been implicated in multiple autoimmune diseases. Strict regulation of IFNβ is critical to minimize uncontrolled inflammation damage to infected hosts while clearing the infection. Together with ATF2/c-Jun, p65/p50, and IRF7, IRF3 form a higher order molecular complex called the IFNβ enhanceosome to strictly regulate IFNβ expression. While the principles of the signaling pathway that activates IRF3 and induces IFNβ expression are established, structural insights into the activation of IRF3 and IFNβ enhanceosome assembly remain scarce. In this proposal, I seek to characterize the structural mechanism of IRF3 activation by using X-ray crystallography. Then, I will use cryo-electron microscopy to characterize the structural features promoting IRF3 interactions with p65 at the enhanceosome. These studies will provide insights into how the other IRFs get activated and how other enhanceosome complexes assemble. Most importantly, structural-based analyses of IRF3 activation and assembly with p65 will provide a basis for designing small molecule modulators that can directly target the activation and suppression of IRF3 in response to viral infections and autoimmune diseases, respectively. !

项目概要 接触病毒后，免疫系统会激活并采取快速措施来保护身体免受病毒侵害 传播感染。我们对导致免疫激活反应的信号级联的理解 病毒感染对于设计调节这一过程的有效治疗药物至关重要。细胞表面和 胞质受体识别病原体特异性分子，例如激活病原体的病毒核酸 干扰素调节因子 3 (IRF3)，是免疫激活的主要调节因子。一旦激活，IRF3 促进干扰素 β (IFNβ) 的表达，这对于抗病毒反应至关重要。尽管IRF3的 IRF3 表达失调与多种自身免疫性疾病有关 疾病。严格调控 IFNβ 对于最大限度地减少受感染宿主不受控制的炎症损害至关重要 同时清除感染。 IRF3 与 ATF2/c-Jun、p65/p50 和 IRF7 一起形成更高阶分子 一种称为 IFNβ 增强体的复合物，可严格调节 IFNβ 的表达。虽然该原则的 激活 IRF3 并诱导 IFNβ 表达的信号通路已建立，结构深入了解 IRF3 和 IFNβ 增强小体组装的激活仍然很少。在这个提案中，我试图描述 使用X射线晶体学研究IRF3激活的结构机制。然后，我将使用冷冻电子 显微镜来表征促进 IRF3 与 p65 在增强体相互作用的结构特征。 这些研究将深入了解其他 IRF 如何被激活以及其他增强体如何 复合物聚集。最重要的是，基于结构的 IRF3 激活和 p65 组装分析 将为设计可直接靶向激活和 抑制 IRF3 分别响应病毒感染和自身免疫性疾病。 ！