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; 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-KB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剂NEMO最近被发现在一种称为Anhidrotic（低糖）外皮异常发育不良的免疫缺陷障碍中突变，具有免疫缺陷（EDA-ID）。 EDA-ID患者易受革兰氏阳性生物的感染，并且由于NEMO中的造成责任突变，因此分子是功能失调的NF-KB信号传导途径。尽管NF-KB途径经过充分研究，以至于它已成为炎症信号转导的范式，但近年来，这种范式已经改变，因为这种途径信号转导强度和配位在关键上取决于赖氨酸-63（K63）连接的多偶联性。最近，NEMO被证明包含一个泛素结合结构域，该结构域对于NF：B信号传导至关重要，而NeMo本身是在两个位点上响应于先天性免疫刺激（NOD2（Crohn的疾病感知性蛋白）和Toll-like受体（TLR）激活的K63-聚泛素化。这种Nemo泛素化对于最佳NF：B信号传导是必需的（1、2、6、13、26）。 EDA-ID中NEMO的点突变的频谱和位置表明，在生化上，它们可能正在干扰Nemo和/或K63连接的Nemo对先天性免疫刺激的泛素结合。此外，EDA-ID中的不同免疫表型和免疫缺陷表明细胞类型的特异性在特定突变方面。最后，该领域很难通过NeMo从NEMO的K63连接的多泛素化中取消泛素结合，因为它与功能相关，最终是NF：B诱导的基因表达。所有这些困难表明，需要系统地评估EDA-ID相关的NEMO突变，涉及NEMO的泛素结合，K63连接的多泛素化和NF-KB相关的基因表达。这些困难还表明，有缺陷的螺旋泛素化的体内模型，以破译其在体内NF：B依赖性基因表达以及炎症和免疫缺陷障碍中的作用。赠款申请旨在回答这些关键问题。 公共卫生相关性：确定对传染病易感性的遗传学的能力导致了新型免疫缺陷形式的鉴定，而挑战是将这些遗传发现转化为疾病的生化机制，以更好地了解该疾病并为该疾病提供更好的治疗。 Nemo是一种对主要炎症信号通路核心的蛋白质，在一种称为EDA-ID的疾病中突变，这种疾病的特征是细菌感染的易感性。该赠款旨在确定Nemo在EDA-ID中的错误，并产生一种可能模仿EDA-ID的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方法。