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

基本信息

批准号：
8672595
负责人：
STEPHEN C BUNNELL
金额：
$ 63.38万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
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/8672595
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

项目摘要

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）：需要适当调节 NF-κB 转录因子来增强对病原体的免疫反应以及免疫系统的正常稳态。相反，过度的 NF-κB 依赖性炎症可能有助于推动自身免疫性疾病的发展和肿瘤的形成。有鉴于此，了解 NF-κB 家族转录因子如何通过影响该通路的不同类型受体的信号传导进行控制非常重要。在抗原激活 T 细胞的过程中，TCR 和 CD28 以及众多信号蛋白被重组为不同的结构，同时经历详细描述的翻译后修饰，例如磷酸化和泛素化。尽管 IKK 复合物在免疫突触中心富含 TCR 的结构域中积累，但受体信号是由接触周围的 TCR“微簇”启动的。我们发现 IKK 复合物被快速招募到含有 TCR 和 Zap70 的信号微簇，但不包括 SLP-76。我们的影像学研究还揭示了额外的 IKK 池？可能涉及 IKK 的交付？ TCR 微簇和/或其下调。然而，关于 IKK 的亚细胞调节，仍有许多重要问题有待解答？及其在 T 细胞激活中的作用。开展此类研究是重要的是，如果我们最终要利用这些知识来操纵 TCR 介导的 NF-κB 激活以达到治疗目的。我们假设经典 NF-κB 通路的 TCR/CD28 激活需要接头蛋白 IKK?/NEMO 的复杂亚细胞调节。因此，在目标 1 中，我们将确定 Zap70 和 Lck 在 IKK 复合物招募至 TCR 微簇中的作用。在目标 2 中，我们将定义 Carma1 和相关蛋白在 IKK 复合物空间调节中的作用。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