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: 3467302
• 负责人: STEPHEN H BENEDICT
• 金额: $ 8.87万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-07-01 至 1993-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3467302
• 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细胞的免疫功能取决于它们的免疫功能。 对抗原作出反应而增殖，从而扩增特异性T细胞 子集 T细胞抗原受体（TCR）的刺激 适当呈递的抗原诱导合成 白细胞介素2（IL 2）受体（IL 2 R）和IL 2。 刺激 IL 2 R与IL 2一起导致T细胞增殖。 我们最近 研究结果表明，虽然早期信号通过这两个 受体不同，一旦信号到达细胞核， 在基因表达上似乎是一样的。 特别是，表达 原癌基因c-fos的表达在两种基因激活后都增加， 受体系统 原癌基因c-fos与许多细胞活化有关 现象，是最早的基因，其合成是诱导 在大多数细胞的激活过程中。 在本建议中，我们将 探讨c-fos调控区参与 激活T细胞。 具体来说，我们将描述约束力 c-fos启动子响应几种配体的活性， 确定响应于每个的DNA结合面积。 我们将 研究是否由各种不同的信号传导途径诱导 刺激在激活c-fos DNA结合蛋白之前会聚 (DBP)，或者单个信号通路是否完全激活 不同的DBP 我们还将研究可能存在或 不存在转录抑制结合蛋白。 我们将 确定在新鲜T中激活的DBP是否存在任何差异 与DBP激活或存在于转化T细胞中相比， 细胞系 我们将决定增加 (Ca2+）i、Na/H反向转运体和两种蛋白激酶PKA和 PKC在c-fos特异性DBP激活中的作用 最后，我们将分离 特异性DBP的抗体，并利用该抗体研究 DBP在静息T细胞中的细胞内定位，以及这是如何 位置可能因激活而改变。 我们将确定 规模、复杂性和潜在的激活或失活 一些c-fos DNA结合复合物的修饰。