LUBAC-dependent regulation of cell death

LUBAC 依赖性细胞死亡调节

• 批准号: 10030736
• 负责人: Jaime Lopez-Mosqueda
• 金额: $ 25.21万
• 依托单位: SOUTH DAKOTA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10030736
• 关键词: Address; Alleles; Animals; Apoptosis; Automobile Driving; CASP1 gene; Cell Death; Cell Death Inhibition; Cell Survival; Cells; Cessation of life; Chronic; Complex; Etiology; Event; Functional disorder; Gene Activation; Genes; Genetic Transcription; Germ-Line Mutation; Goals; Immune; In Vitro; Inflammation; Inflammatory; Lead; Life; Mediating; Modification; Molecular; Mus; NF-kappa B; Organ; Outcome; Pathway interactions; Patients; Phenocopy; Phenotype; Phosphorylation; Post-Translational Protein Processing; Prevention; Production; Proteins; Publishing; Regulation; Research; Role; Sepsis; Signal Transduction; Signaling Molecule; Site; TNF gene; Testing; Tissues; Translations; Tumor Necrosis Factor Receptor; Ubiquitin; Ubiquitination; Work; base; cohort; congenital immunodeficiency; cytokine; in vivo; insight; mouse model; mutant; prevent; response; sharpin; systemic inflammatory response; tool; ubiquitin ligase; ubiquitin-protein ligase

PROJECT SUMMARY Tumor necrosis factor (TNF) is an important cytokine that coordinates cytokine production, inflammation, cell survival, and cell death. How life and death decisions are made in response to TNF is not completely understood. Several molecular events determine whether TNF stimulation of cells leads to a transcriptional response that promotes cell survival or whether it can lead to cell death. Signaling downstream of the TNF receptor is heavily regulated by post-translation modifications including ubiquitination. LUBAC, a ubiquitin (Ub) E3 ligase consisting of Sharpin, HOIL and HOIP, is the only Ub ligase described to date that can modify proteins with linear Ub chains (Met1-Ub chains). LUBAC activity is essential for NF-kB dependent transcription of prosurvival genes upon TNF stimulation. Cells derived from patients with germline mutations in LUBAC components elicit defective NF-kB- dependent gene transcription and aberrant activation of cell death pathways. Similarly, murine models with defective LUBAC components elicit chronic inflammation and increased apoptosis in multiple organs and tissues. Our published studies indicate that cell death, rather than a deficiency in NF-kB dependent gene transcription, in LUBAC-deficient (sharpincpdm) animals is driving the TNF-dependent chronic inflammation as this phenotype is reversed by a compound deficiency in FADD—an important component of the death inducing signaling complex. This insight led us to the hypothesis that LUBAC activity is directly required for the prevention of cell death. In aim 1, we will address the molecular basis for LUBAC modification of protein targets with Met1-Ub chains. We will address how LUBAC can modify FADD with Met1-Ub chains. In aim 2, we will characterize the functional relevance of FADD modification in cells and use a new Ub-based tool to enrich for Met1-Ub chains. In aim 3, we will investigate the function of LUBAC auto-ubiquitination and how phosphorylation can regulate LUBAC’s linear Ub chain forming activity. Our studies will provide mechanistic insight into the etiology of primary immune deficiencies associated with patients with germline mutations in LUBAC components.

项目概要 肿瘤坏死因子（TNF）是一种重要的细胞因子，协调细胞因子的产生、炎症、细胞 生存和细胞死亡。如何根据 TNF 做出生死决定尚不完全清楚。 几个分子事件决定 TNF 对细胞的刺激是否会导致转录反应 促进细胞存活或是否会导致细胞死亡。 TNF 受体的下游信号传导非常重要 受翻译后修饰（包括泛素化）调节。 LUBAC，一种泛素 (Ub) E3 连接酶，由 Sharpin、HOIL 和 HOIP 的 Ub 连接酶是迄今为止描述的唯一可以用线性 Ub 链修饰蛋白质的 Ub 连接酶 （Met1-Ub 链）。 LUBAC 活性对于 TNF 上的促生存基因的 NF-kB 依赖性转录至关重要 刺激。来自 LUBAC 成分种系突变患者的细胞会引发有缺陷的 NF-kB- 依赖性基因转录和细胞死亡途径的异常激活。同样，小鼠模型 有缺陷的 LUBAC 成分会引发慢性炎症并增加多个器官和组织的细胞凋亡。 我们发表的研究表明细胞死亡，而不是 NF-kB 依赖性基因转录缺陷， 在 LUBAC 缺陷 (sharpincpdm) 动物中，这种表型正在驱动 TNF 依赖性慢性炎症 FADD（死亡诱导信号的重要组成部分）中的化合物缺陷可逆转这种情况 复杂的。这一见解使我们得出这样的假设：LUBAC 活性是预防细胞损伤直接所必需的。 死亡。在目标 1 中，我们将研究利用 Met1-Ub 修饰蛋白质靶标的 LUBAC 的分子基础 链。我们将讨论 LUBAC 如何使用 Met1-Ub 链修改 FADD。在目标 2 中，我们将描述 细胞中 FADD 修饰的功能相关性，并使用基于 Ub 的新工具来富集 Met1-Ub 链。在 目标3，我们将研究LUBAC自动泛素化的功能以及磷酸化如何调节 LUBAC 的线性 Ub 链形成活性。我们的研究将为原发性疾病的病因学提供机制上的见解 与 LUBAC 成分种系突变的患者相关的免疫缺陷。