Phosphorylation mimetic motif of cryptochrome proteins blocks IRF3 activation

隐花色素蛋白的磷酸化模拟基序可阻断 IRF3 激活

• 批准号: 10583785
• 负责人: Shitao Li
• 金额: $ 22.85万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-11-15 至 2024-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10583785
• 关键词: Adaptor Signaling Protein; Alanine; Asthma; Attenuated; Autoimmune Diseases; Binding; C-terminal; Circadian Rhythms; Communicable Diseases; Consensus; Data; Dimerization; Docking; Exhibits; Genes; Glutamic Acid; Goals; Host Defense; Human; IRF3 gene; Immune; Immune System Diseases; Influenza A virus; Innate Immune Response; Innate Immune System; Interferon Type I; Knock-out; Knowledge; Literature; Mediating; Molecular; Mutation; Natural Immunity; Nucleic Acids; Pathology; Pattern; Pattern recognition receptor; Periodicity; Phosphorylation; Phosphorylation Site; Pilot Projects; Polyubiquitin; Production; Proteins; Proteomics; Rheumatoid Arthritis; Role; Signal Pathway; Signal Transduction; Site; Stimulator of Interferon Genes; Symptoms; TBK1 gene; TLR3 gene; TRAF2 gene; Testing; Tretinoin; Ubiquitination; Virus Diseases; autoinflammatory diseases; circadian; cryptochrome; cryptochrome 2; human disease; hydrophilicity; in vivo; mimetics; novel; novel therapeutics; pathogen; recruit; Adaptor Signaling Protein; Alanine; Asthma; Attenuated; Autoimmune Diseases; Binding; C-terminal; Circadian Rhythms; Communicable Diseases; Consensus; Data; Dimerization; Docking; Exhibits; Genes; Glutamic Acid; Goals; Host Defense; Human; IRF3 gene; Immune; Immune System Diseases; Influenza A virus; Innate Immune Response; Innate Immune System; Interferon Type I; Knock-out; Knowledge; Literature; Mediating; Molecular; Mutation; Natural Immunity; Nucleic Acids; Pathology; Pattern; Pattern recognition receptor; Periodicity; Phosphorylation; Phosphorylation Site; Pilot Projects; Polyubiquitin; Production; Proteins; Proteomics; Rheumatoid Arthritis; Role; Signal Pathway; Signal Transduction; Site; Stimulator of Interferon Genes; Symptoms; TBK1 gene; TLR3 gene; TRAF2 gene; Testing; Tretinoin; Ubiquitination; Virus Diseases; autoinflammatory diseases; circadian; cryptochrome; cryptochrome 2; human disease; hydrophilicity; in vivo; mimetics; novel; novel therapeutics; pathogen; recruit

Project summary The innate immune system employs germline-encoded pattern recognition receptors (PRRs) to detect common pathogen-associated molecular patterns. However, only several PRR signaling pathways, including RIG-I- MAVS, cGAS-STING, and TLR3/4-TRIF, activate interferon regulatory factor 3 (IRF3). Recent studies showed that the innate immune adaptors (MAVS, STING, and TRIF) are phosphorylated at their respective C-terminal consensus motif, pLxIS (p, hydrophilic residue; x, any residue; S, phosphorylation site) by TBK1. The phosphorylation of the pLxIS motif in innate immune adaptor proteins is an essential and conserved mechanism that selectively recruits IRF3 to activate type I IFN production. However, whether and how the host or pathogen utilizes this motif to attenuate or block IRF3 signaling is unknown. Our overall objective in this application is to determine the role of the cryptochrome (CRY) proteins in IRF3 signaling pathways and host defense to viral infection. Our preliminary proteomic data showed that cryptochrome (CRY) proteins, CRY1 and CRY2, interacted with IRF3. Interestingly, CRY1 and CRY2 have two and one conserved pLxIE motifs, respectively. As glutamic acid (E) is a phosphorylation mimic, this pLxIE motif might be a constitutively phosphorylated pLxIS. Our pilot studies showed that mutation of E to alanine (A) in the pLxIE abolished the binding to IRF3, suggesting that the pLxIE motif is also a docking site for IRF3. Furthermore, deficiency of both CRY proteins also augmented IRF3 signaling and type I IFN production. However, how CRY proteins inhibit innate immune responses is not well elucidated. Based on the existing literature and our preliminary data, we hypothesize that CRY proteins are decoy adaptors for IRF3 via the pLxIE motif and the binding blocks interaction between IRF3 and innate immune adaptors, thereby inhibiting IRF3 phosphorylation and activation. Aim 1 will determine the role of CRY1/2 in the RIG-I-mediated IRF3 signaling pathway. Aim 2 will dissect the molecular mechanisms of how CRY1/2 suppresses IRF3-mediated innate immunity. Many human immune diseases exhibit circadian rhythmicity in their symptoms and pathology, including asthma, rheumatoid arthritis, and other disorders of the immune system. Our study will bridge the gaps in understanding the role of the circadian proteins CRY1/2 in host innate immune response. This novel mechanism proposed in our application will advance our understanding of the crosstalk between innate immunity and circadian.

项目摘要 先天免疫系统采用种系编码的模式识别受体（PRR）来检测常见 病原体相关的分子模式。但是，只有几个PRR信号通路，包括rig-i- MAVS，CGAS-STING和TLR3/4-TRIF激活干扰素调节因子3（IRF3）。最近的研究表明 先天免疫适配器（MAV，STING和TRIF）在其各自的C末端被磷酸化 共识基序，PLXI（P，亲水性残基； X，任何残基； S，磷酸化位点）。这 先天免疫适配器蛋白中PLXIS基序的磷酸化是必不可少的和保守的 有选择地募集IRF3激活I型IFN产生的机制。但是，是否以及如何主持人 或病原体利用此基序来减轻或阻止IRF3信号传导。我们的整体目标 应用是确定加密块状（CRY）蛋白在IRF3信号通路和主机中的作用 对病毒感染的防御。我们的初步蛋白质组学数据表明，隐性（CRY）蛋白质Cry1 和Cry2，与IRF3相互作用。有趣的是，Cry1和Cry2具有两个和一个保守的plxie图案， 分别。由于谷氨酸（E）是一种磷酸化模拟物，因此该plxie基序可能是组成型的 磷酸化的PLXI。我们的试点研究表明，plxie中E向丙氨酸的突变废除了 与IRF3的结合，表明PLXIE基序也是IRF3的对接位点。此外，两者的不足 哭泣蛋白还增强了IRF3信号传导和I型IFN产生。但是，哭泣蛋白如何抑制 先天免疫反应未得到很好的阐明。根据现有文献和我们的初步数据，我们 假设哭泣蛋白是通过PLXIE基序和结合块的IRF3的诱饵适配器 IRF3与先天免疫适配器之间的相互作用，从而抑制IRF3磷酸化和激活。 AIM 1将确定CRY1/2在RIG-I介导的IRF3信号通路中的作用。 AIM 2将剖析 CRY1/2如何抑制IRF3介导的先天免疫的分子机制。 许多人类免疫疾病在其症状和病理学中表现出昼夜节律，包括 哮喘，类风湿关节炎和免疫系统的其他疾病。我们的研究将弥合差距 了解昼夜节律蛋白Cry1/2在宿主先天免疫反应中的作用。这本小说 我们应用中提出的机制将提高我们对先天串扰的理解 免疫力和昼夜节律。