GTP-BINDING PROTEIN RHO IN T CELL SIGNALING PATHWAYS

T 细胞信号通路中的 GTP 结合蛋白 RHO

• 批准号: 2671469
• 负责人: JONATHAN P MOORMAN
• 金额: $ 8.78万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-07-01 至 2001-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2671469
• 关键词: T cell receptor; T lymphocyte; biological signal transduction; fluorimetry; guanine nucleotide binding protein; immunoprecipitation; interleukin 2; leukocyte activation /transformation; polymerase chain reaction; protein kinase C; tissue /cell culture

Having been clinically trained in Internal Medicine and Infectious Diseases. I have long been intrigued by the biological responses to disease. My interest in Infectious Diseases and the basic processes that govern immune responses led me to the study of signal transduction pathways in leukocytes. Over the last several years. I have devoted the majority of my time to developing an understanding of the molecular biology and biochemistry of lymphocyte signalling and particularly the importance of small G proteins in this process. Building upon this foundation, my goal is to make the transition from a postdoctoral researcher to an independent investigator with an expertise in basic signal transduction in leukocytes. The T cell receptor complex (TCR) mediated signalling processes that are necessary for activation and proliferation of lymphocytes during immune responses. Ligation of the TCR by antigenic stimuli drives cellular responses that include upregulation of cytokine gene expression, and, ultimately, T cell activation. In addition, ligation of the TCR leads to remodeling of the action cytoskeleton; interactions between the TCR and this action cytoskeleton are crucial for effective T cell activation. Members of the Rho subfamily of GTPases are likely candidates to function as regulators of TCR-mediated signal transduction pathways. The role of Rho in lymphocytes, however, has received little attention. Studies in other systems suggest that Rho likely coordinates intracellular signalling pathways regulating both cytoskeletal remodeling and protein kinase-activated gene expression. The overall objective of this work is to investigate the hypothesis that Rho proteins coordinate such dual signal transduction pathways in lymphocytes, specifically those pathways emerging from TCR engagement and leading to effective T cell activation. Our study aims will be to 1) establish the role of Rho during classic T cell activation pathways, and to 2) characterize the function of Rho in the interaction between the TCR and early signalling events occurring at the plasma membrane. Using a transient expression system based on Sindbis virus we will express Rho proteins, including constitutively active and dominant negative mutants. Inlyphocytes; in addition, we will express C3 exoenzyme a specific inhibitor of Rho function. These tools will allow us to carefully delineate the role of Rho during T cell activation as assessed by both downstream cytokine gene expression and by alterations in early TCR signalling events at the plasma membrane. The environment in which I will complete this project is ideal. Experts in the areas of immunology. G protein signalling and protein biochemistry are committed to my efforts and will provide a rigorous academic milieu in which I can develop my skills effectively. Advocates within the Beirne Carter Center for Immunology and the Departments of Medicine and Microbiology will insure that I receive excellent training as I make the transition to an independent investigator.

接受过内科和感染学的临床培训 疾病。长期以来，我一直对生物反应很感兴趣 为了疾病。我对传染病和基础知识的兴趣 支配免疫反应的过程使我研究了 白细胞中的信号转导途径。在过去的几年里 好几年了。我把我的大部分时间都花在开发一个 对细菌的分子生物学和生物化学的认识 淋巴细胞信号传递，特别是小G的重要性 在这一过程中的蛋白质。在此基础上，我的目标是 从博士后研究员过渡到 具有基本信号专业知识的独立调查员 在白细胞中的转导。T细胞受体复合体(TCR) 激活和激活所需的中介信号传递过程 免疫反应过程中淋巴细胞的增殖。结扎 通过抗原刺激产生的TCR驱动细胞反应， 包括上调细胞因子基因表达，最终， T细胞活化。此外，TCR的结扎导致 作用细胞骨架的重塑； TCR和这种作用的细胞骨架对有效的T细胞至关重要 激活。GTP酶的Rho亚家族成员很可能 作为TCR介导的信号调节器的候选人 转导通路。然而，Rho在淋巴细胞中的作用， 几乎没有受到关注。对其他系统的研究表明 Rho可能协调细胞内信号通路调节 细胞骨架重塑与蛋白激酶激活基因 表情。这项工作的总体目标是调查 Rho蛋白协调这种双重信号的假说 淋巴细胞中的转导通路，特别是那些通路 从TCR参与中脱颖而出并产生有效的T细胞 激活。我们的研究目的将是：1)确定RHO的角色 在经典的T细胞激活途径中，并2)表征 Rho在TCR与EARE相互作用中的作用 发生在质膜上的信号事件。使用 我们将要表达的基于Sindbis病毒的瞬时表达系统 Rho蛋白，包括结构活性蛋白和优势蛋白 阴性变种人。在淋巴细胞中；此外，我们还将表达C3 胞外酶是Rho功能的特异性抑制物。这些工具将 允许我们仔细描述Rho在T细胞中的作用 下游两种细胞因子基因评估的激活 表达和早期TCR信号事件的变化 质膜。我将完成这项工作的环境 项目是理想的。免疫学领域的专家。G蛋白 信号和蛋白质生物化学致力于我的努力 并将提供一个严谨的学术环境，在其中我可以 有效地发展我的技能。贝恩·卡特内部的倡导者 免疫学中心和医学系 微生物学将确保我在制作过程中得到出色的训练 向独立调查员过渡。