REGULATION OF C-FOS EXPRESSION DURING T CELL ACTIVATION

T 细胞激活过程中 C-FOS 表达的调节

• 批准号: 3467305
• 负责人: STEPHEN H BENEDICT
• 金额: $ 11.97万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-09-01 至 1993-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3467305
• 关键词: DNA binding protein; T cell receptor; T lymphocyte; affinity chromatography; antibody; antiport; calcium; cytokine receptors; gene expression; genetic manipulation; genetic promoter element; genetic transcription; immunofluorescence technique; interleukin 2; laboratory rabbit; leukocyte activation /transformation; nucleic acid sequence; protein kinase; protein kinase C; protooncogene

The immune function of T cells is dependent on their ability to proliferate in response to antigen, thus expanding specific T cell subsets. Stimulation of the T cell antigen receptor (TCR) with appropriately presented antigen induces synthesis of the interleukin 2 (IL2) receptor (IL2R), and of IL2. Stimulation of the IL2R with IL2 causes the T cells to proliferate. Our recent findings indicate that although early signaling through these two receptors differ, once a signal reaches the nucleus, early changes in gene expression seem to be the same. In particular, expression of the proto-oncogene c-fos is increased after activation of both receptor systems. The proto-oncogene c-fos is associated with many cell activation phenomena, and is the earliest gene whose synthesis is induced during activation of most cells. In the present proposal, we will explore the participation of the c-fos regulatory region in the activation of T cells. Specifically, we will delineate the binding activity of the c-fos promoter in response to several ligands and determine the area of DNA bound in response to each. We will investigate whether signaling pathways induced by the various stimuli converge before activating c-fos DNA binding proteins (DBP), or whether individual signaling pathways activate entirely different DBPs. We will also examine the possible existence or nonexistence of transcription-inhibiting binding proteins. We will determine if any differences exist in the DBP activated in fresh T cells compared with DBP activated or present in transformed T cell lines. We will determine the participation of increased (Ca2+)i, the Na/H antiport and the two protein kinases PKA and PKC in activation of c-fos specific DBP. Finally, we will isolate antibody against specific DBP and use this anitbody for study of intracellular location of the DBP in resting T cells, and how this location may change as a result of activation. We will determine the size, complexity, and potential activating or inactivating modifications of some of the c-fos DNA binding complexes.

T细胞的免疫功能依赖于它们的能力 对抗原作出反应而增殖，从而扩增特定的T细胞 子集。对T细胞抗原受体(TCR)的刺激 适当提呈的抗原诱导合成 白介素2(IL2)受体(IL2R)和IL2。对人的刺激 IL-2R和IL-2可导致T细胞增殖。我们最近 研究结果表明，尽管通过这两个基因发出的早期信号 受体不同，一旦信号到达细胞核，早期就会发生变化 在基因表达上似乎是一样的。尤其是，表达式 原癌基因c-fos在两者激活后表达增加 受体系统。 原癌基因c-fos与许多细胞激活有关 现象，是最早被诱导合成的基因 在大多数细胞激活期间。在目前的建议中，我们将 探索c-fos监管区域参与 激活T细胞。具体地说，我们将描述绑定 C-fos启动子对几种配体和 确定与每个DNA结合的区域。我们会 调查各种不同的信号传导途径是否 刺激在激活c-fos DNA结合蛋白之前收敛 (DBP)，或者单个信号通路是否完全激活 不同的数据库管理员。我们还将检查可能存在的或 不存在转录抑制结合蛋白。我们会 确定在Fresh T中激活的DBP是否存在任何差异 转化T细胞与DBP激活或存在的细胞比较 细胞系。我们将确定增加的参与 (Ca~(2+))i、Na/H逆向转运和两个蛋白激酶PKA和PKA PKC在c-fos特异性DBP激活中的作用。最后，我们将分离出 特异性DBP抗体的制备及应用研究 DBP在静息T细胞中的细胞内定位 位置可能会因激活而改变。我们将决定 大小、复杂性和潜在的激活或停用 一些c-fos DNA结合复合体的修饰。