The Role of NEMO Ubiquitination in EDA-ID

NEMO 泛素化在 EDA-ID 中的作用

• 批准号: 8227941
• 负责人: Derek W Abbott
• 金额: $ 19.63万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-11 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8227941
• 关键词: Acetylation; Amides; Applications Grants; Area; B-Lymphocytes; Bacterial Infections; Binding; Binding Proteins; Biochemical; Biochemistry; Bloch Sulzberger syndrome; CD4 Positive T Lymphocytes; Cells; Characteristics; Circular Dichroism; Clinical Pathways; Co-Immunoprecipitations; Coiled-Coil Domain; Communicable Diseases; Crohn' s disease; Disease; Disease susceptibility; Dissociation; Distant; Ectodermal Dysplasia; Event; Female; Gene Activation; Gene Expression; Genes; Genetic; Genetic Predisposition to Disease; Gram-Positive Bacterial Infections; Grant; Helper-Inducer T-Lymphocyte; Heterozygote; Immune; Immunologic Deficiency Syndromes; Immunologics; Infection; Inflammatory; Lead; Link; Location; Lysine; Manuscripts; Missense Mutation; Mus; Mutate; Mutation; NF-kappa B; NFKB Signaling Pathway; Organism; Pathology; Pathway interactions; Patients; Peptides; Phenocopy; Phenotype; Play; Point Mutation; Polyubiquitin; Polyubiquitination; Predisposition; Process; Proteins; Receptor Activation; Role; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Site; Skin Abnormalities; Specificity; Stimulus; Structure; Surface; System; T-Lymphocyte; Testing; Toll-like receptors; Transduction Gene; Translating; Ubiquitin; Ubiquitination; Work; X Chromosome; X Inactivation; cell type; in vivo; in vivo Model; macrophage; male; mouse model; mutant; novel; public health relevance; reconstitution; response; skeletal abnormality

DESCRIPTION (provided by applicant): NF-kB signaling lies at the center of Immunodeficiency and Inflammatory diseases. A key regulator of NF-kB signal transduction, NEMO, has recently been found to be mutated in an Immunodeficiency Disorder called called Anhidrotic (hypohydrotic) Ectodermal Dysplasia with Immunodeficiency (EDA-ID). Patients with EDA-ID are susceptible to infection with gram-positive organisms, and molecularly, a dysfunctional NF-kB signaling pathway, owing to hypomorphic mutations in nemo is to blame. While the NF-kB pathway is so well-studied that it has become a paradigm for inflammatory signal transduction, this paradigm has shifted in recent years with the recognition that this pathways signal transduction strength and coordination is critically dependent on Lysine-63 (K63)-linked polyubiquitination. NEMO was very recently shown to contain a ubiquitin binding domain that is essential for NF:B signaling, and NEMO, itself, is K63-polyubiquitinated on two sites in response to innate immune stimulation (NOD2 (Crohn's Disease-susceptibility protein) and Toll-like Receptor (TLR) activation). This NEMO ubiquitination is necessary for optimal NF:B signaling (1, 2, 6, 13, 26). The spectrum and location of point mutations of nemo in EDA-ID suggest that biochemically, they may be interfering with ubiquitin binding by NEMO and/or K63-linked polyubiquitination of NEMO in response to innate immune stimuli. In addition, the varied immunologic phenotypes and immunodeficiencies in EDA-ID suggest cell-type specificity in regards to the particular mutations. Lastly, it has been difficult for the field to uncouple ubiquitin binding by NEMO from K63-linked polyubiquitination of NEMO as it relates to function and ultimately, NF:B-induced gene expression. All these difficulties point to the need to systematically evaluate EDA-ID- associated NEMO mutations in regards to ubiquitin binding, K63-linked polyubiquitination of NEMO and NF-kB-associated gene expression. These difficulties also point to a need for an in vivo model of defective NEMO ubiquitination to decipher its role in in vivo NF:B-dependent gene expression and inflammatory and immunodeficiency disorders. The grant application aims to answer these key questions. PUBLIC HEALTH RELEVANCE: The ability to determine the genetics of susceptibility to infectious disease has led to the identification of novel forms of immunodeficiency, and the challenge is to translate these genetic findings into biochemical mechanisms of disease to both better understand the disease and allow better treatment for that disease. NEMO, a protein that is central to the major inflammatory signaling pathway is mutated in a disease called EDA-ID, a disease that is characterized by susceptibility to bacterial infections. This grant aims to determine how NEMO is faulty in EDA-ID and to generate a mouse model that might mimic many of the features of EDA-ID in hopes that this might lead to a better understanding of and better treatments for this disease.

描述（由申请人提供）：NF-κ B信号传导位于免疫缺陷和炎症性疾病的中心。NF-kB信号转导的关键调节因子NEMO最近被发现在称为无汗性（少汗性）外胚层发育不良伴免疫缺陷（EDA-ID）的免疫缺陷疾病中发生突变。EDA-ID患者易感染革兰氏阳性菌，分子上，NF-kB信号通路功能障碍，由于nemo的亚型突变是罪魁祸首。虽然NF-kB通路被充分研究，它已成为炎症信号转导的范例，但近年来随着认识到该通路的信号转导强度和协调严重依赖于赖氨酸-63（K63）连接的多聚泛素化，这种范例已经发生了变化。NEMO最近被证明含有NF：B信号传导所必需的泛素结合结构域，并且NEMO本身在响应先天免疫刺激（NOD 2（克罗恩病易感蛋白）和Toll样受体（TLR）激活）的两个位点上被K63-聚泛素化。这种NEMO泛素化是最佳NF：B信号传导所必需的（1，2，6，13，26）。EDA-ID中NEMO点突变的谱和位置表明，在生物化学上，它们可能干扰NEMO的泛素结合和/或响应于先天免疫刺激的NEMO的K63连接的多泛素化。此外，EDA-ID中不同的免疫表型和免疫缺陷表明特定突变的细胞类型特异性。最后，本领域难以将NEMO的泛素结合与NEMO的K63连接的多聚泛素化解偶联，因为它涉及功能并最终涉及NF：B诱导的基因表达。所有这些困难都表明需要系统地评估EDA-ID相关NEMO突变的泛素结合，K63连接的NEMO多聚泛素化和NF-κ B相关基因表达。这些困难也指出需要一个体内模型的缺陷NEMO泛素化，以破译其在体内NF：B依赖性基因表达和炎症和免疫缺陷疾病的作用。申请补助金的目的是回答这些关键问题。 公共卫生相关性：确定感染性疾病易感性的遗传学的能力导致了免疫缺陷的新形式的鉴定，挑战是将这些遗传学发现转化为疾病的生化机制，以更好地理解疾病并更好地治疗该疾病。NEMO是一种蛋白质，是主要炎症信号通路的核心，在一种称为EDA-ID的疾病中发生突变，这种疾病的特征是对细菌感染的易感性。这项资助旨在确定NEMO在EDA-ID中的缺陷，并生成一个可能模仿EDA-ID许多特征的小鼠模型，希望这可能会导致对这种疾病的更好理解和更好的治疗。