Identification of a novel cyclosporin-sensitive element in the human tumor necrosis factor alpha gene promoter.
Identification of a novel cyclosporin-sensitive element in the human tumor necrosis factor alpha gene promoter.
复制标题
人肿瘤坏死因子α基因启动子中新型环孢菌素敏感元件的鉴定。
DOI：
10.1084/jem.178.4.1365
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发表时间：
1993-10-01
影响因子：
15.3
通讯作者：
Rao, A
中科院分区：
文献类型：
--
作者：
Goldfeld, A E;McCaffrey, P G;Strominger, J L;Rao, A
肿瘤坏死因子α（TNF-α）是一种具有多效性生物效应的细胞因子，由多种细胞类型响应不同刺激的诱导而产生。在本文中，通过 T 细胞受体 (TCR) 配体或单独的钙离子载体刺激，TNF-α mRNA 在小鼠 T 细胞克隆中被高度诱导。诱导快速，不需要从头合成蛋白质，并且被免疫抑制剂环孢菌素 A (CsA) 完全阻断。我们已经鉴定出人类 TNF-α 启动子元件 kappa 3，它在该基因的钙介导诱导性和 CsA 敏感性中发挥关键作用。在电泳迁移率变动分析中，含有 kappa 3 的寡核苷酸与未刺激 T 细胞提取物中存在的蛋白质形成两个 DNA 蛋白质复合物。这些复合物仅在 T 细胞刺激后才出现在核提取物中。可诱导核复合物的诱导是快速的，不依赖于蛋白质合成，并且被 CsA 阻断，因此，与 TCR 配体或钙离子载体对 TNF-α mRNA 的诱导完全平行。我们的研究表明 kappa 3 结合因子类似于活化 T 细胞核因子中预先存在的成分。因此，TNF-α基因是活化T细胞中的立即早期基因，并提供了研究活化T细胞中CsA敏感基因诱导的新模型系统。
Tumor necrosis factor alpha (TNF-alpha), a cytokine with pleiotropic biological effects, is produced by a variety of cell types in response to induction by diverse stimuli. In this paper, TNF-alpha mRNA is shown to be highly induced in a murine T cell clone by stimulation with T cell receptor (TCR) ligands or by calcium ionophores alone. Induction is rapid, does not require de novo protein synthesis, and is completely blocked by the immunosuppressant cyclosporin A (CsA). We have identified a human TNF-alpha promoter element, kappa 3, which plays a key role in the calcium-mediated inducibility and CsA sensitivity of the gene. In electrophoretic mobility shift assays, an oligonucleotide containing kappa 3 forms two DNA protein complexes with proteins that are present in extracts from unstimulated T cells. These complexes appear in nuclear extracts only after T cell stimulation. Induction of the inducible nuclear complexes is rapid, independent of protein synthesis, and blocked by CsA, and thus, exactly parallels the induction of TNF-alpha mRNA by TCR ligands or by calcium ionophore. Our studies indicate that the kappa 3 binding factor resembles the preexisting component of nuclear factor of activated T cells. Thus, the TNF-alpha gene is an immediate early gene in activated T cells and provides a new model system in which to study CsA-sensitive gene induction in activated T cells.