Biochemical and Spatial Regulation of IKKg/NEMO During T Cell Activation

T 细胞激活过程中 IKKg/NEMO 的生化和空间调节

• 批准号: 8582089
• 负责人: STEPHEN C BUNNELL
• 金额: $ 55万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-06-06 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8582089
• 关键词: Adaptor Signaling Protein; Address; Alleles; Anti-Inflammatory Agents; Anti-inflammatory; Antigen Receptors; Antigens; Autoimmune Diseases; B-Lymphocytes; Biochemical; Biological; Biological Assay; CD27 Antigens; CD28 gene; Cells; Cellular Structures; Chimera organism; Chronic; Complex; Data; Development; Down-Regulation; Elements; Event; Family; Fluorescence Resonance Energy Transfer; Health; Homeostasis; Image; Immune; Immune response; Immune system; Individual; Inflammation; Inflammatory; Knowledge; LCP2 gene; Laboratories; Lead; Light; Malignant Neoplasms; Maps; Mediating; Nature; Pathway interactions; Phosphorylation; Play; Polyubiquitin; Post-Translational Protein Processing; Protein Family; Protein Tyrosine Kinase; Proteins; Receptor Signaling; Recruitment Activity; Regulation; Relative (related person); Reporter; Role; Series; Signal Pathway; Signal Transduction; Signaling Protein; Structure; Synapses; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Therapeutic; Ubiquitination; base; cell growth regulation; cell type; conserved helix-loop-helix ubiquitous kinase; inhibitor/antagonist; mutant; novel; pathogen; protein complex; receptor; scaffold; spatial relationship; spatiotemporal; transcription factor; transmission process; tumor

DESCRIPTION (provided by applicant): Proper regulation of NF-?B transcription factors is required to mount immune responses to pathogens and for the normal homeostasis of the immune system. Conversely, excessive NF- ?B-dependent inflammation can help to drive the development of autoimmune disease and the formation of tumors. In this light, it is important to know how NF-?B family transcription factors are controlled by signaling through the different types of receptors that impinge on this pathway. During T cell activation by antigen, TCR and CD28, and numerous signaling proteins, are reorganized into distinct structures while undergoing well-described post-translational modifications, such as phosphorylation and ubiquitination. Although the IKK complex accumulates in the TCR-rich domain at the center of the immune synapse, receptor signals are initiated by TCR 'microclusters' in the periphery of the contact. We have found that the IKK complex is rapidly recruited to signaling microclusters containing the TCR and Zap70, but not SLP-76. Our imaging studies have also revealed additional pools of IKK? that may be involved in delivery of IKK? to TCR microclusters and/or its down-regulation. However, a number of important questions remain to be answered regarding the sub-cellular regulation of IKK? and its role in T cell activation. Carrying out such studies is important, if we are to eventually exploit this knowledge to manipulate TCR-mediated NF-?B activation for therapeutic purposes. We hypothesize that TCR/CD28 activation of the classical NF-?B pathway requires complex sub-cellular regulation of the adaptor protein IKK?/NEMO. Thus, in Aim 1, we will determine the role of Zap70 and Lck in recruitment of the IKK complex to TCR microclusters. In Aim 2, we will define the role of Carma1 and associated proteins in spatial regulation of the IKK complex. Finally, in Aim 3 we will define the sub-cellular compartment(s) containing total and active IKK complexes

描述（由申请人提供）：NF-？B转录因子是启动对病原体的免疫应答和维持免疫系统正常稳态所必需的。相反，过量的NF-？B依赖性炎症可以帮助驱动自身免疫性疾病的发展和肿瘤的形成。在这种情况下，重要的是要知道如何NF-？B家族转录因子通过影响该途径的不同类型受体的信号传导来控制。在T细胞被抗原活化期间，TCR和CD 28以及许多信号传导蛋白被重组成不同的结构，同时经历充分描述的翻译后修饰，如磷酸化和泛素化。尽管IKK复合物在免疫突触中心的富含TCR的结构域中积累，但受体信号由接触周边的TCR“微簇”启动。我们已经发现IKK复合物被迅速募集到含有TCR和Zap 70的信号微簇中，而不是SLP-76。我们的影像学研究还显示了IKK？可能参与IKK的传递TCR微簇和/或其下调。然而，一些重要的问题仍然有待回答的亚细胞调控IKK？及其在T细胞活化中的作用。开展这些研究是 重要的是，如果我们最终利用这些知识来操纵TCR介导的NF-？B激活用于治疗目的。我们假设TCR/CD 28激活经典NF-？B途径需要接头蛋白IKK？/ NEMO因此，在目的1中，我们将确定Zap 70和Lck在IKK复合物募集到TCR微簇中的作用。在目标2中，我们将确定的作用Carma 1和相关蛋白的IKK复合物的空间调节。最后，在目标3中，我们将定义包含总IKK复合物和活性IKK复合物的亚细胞区室