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首页> 外文期刊>The Journal of Immunology: Official Journal of the American Association of Immunologists >An activation-responsive element in single C motif-1/lymphotactin promoter is a site of constitutive and inducible DNA-protein interactions involving nuclear factor of activated T cell.
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An activation-responsive element in single C motif-1/lymphotactin promoter is a site of constitutive and inducible DNA-protein interactions involving nuclear factor of activated T cell.

机译：单个C基元1 /淋巴细胞生成素启动子中的活化响应元件是涉及活化T细胞核因子的组成型和诱导型DNA-蛋白质相互作用的位点。

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摘要

Single C motif-1 (SCM-1)/lymphotactin is a C-type chemokine whose expression is activation dependent, cyclosporin A sensitive and restricted to CD8+ T cells, double-negative thymocytes, gammadelta-type T cells, and NK cells. In humans, there are two highly homologous genes encoding SCM-1alpha and SCM-1beta. Here we examined the regulatory mechanism of the SCM-1 genes. The luciferase reporter gene under the control of the 5' flanking region of 0.7 kb was strongly induced upon activation with anti-CD3 or PHA plus PMA only in SCM-1-producer T cell lines through a cyclosporin A-sensitive mechanism. An element termed E1 located at -108 to -95 nt relative to the major transcription start site was found to be critical for the promoter activity. In electrophoretic mobility shift assays using the E1 oligonucleotide as probe, nuclear extracts from unstimulated T and B cell lines formed a constitutive complex termed complex I, while nuclear extracts from stimulated SCM-1-producer T cell lines formed a higher mobility complex termed complex II with a concomitant decrease in complex I. The shift from complex I to complex II seen only in SCM-1-producer T cell lines upon activation was completely suppressed by cyclosporin A. Both complexes were critically dependent on the NF-AT core sequence TTTCC in the E1 element and were partially supershifted by anti-NF-ATp. One-hybrid assays in yeast isolated NF-ATp as an E1 binding protein, and transfection of NF-ATp into T and B cell lines strongly enhanced the activation-dependent SCM-1 promoter activity. Collectively, a unique mechanism involving NF-ATp appears to regulate the cell type-specific and activation-dependent expression of the SCM-1 genes.
机译：单个C基序1（SCM-1）/淋巴动蛋白是一种C型趋化因子，其表达依赖于激活，环孢菌素A敏感，并仅限于CD8 + T细胞，双阴性胸腺细胞，γ型T细胞和NK细胞。在人类中，有两个高度同源的基因编码SCM-1alpha和SCM-1beta。在这里，我们检查了SCM-1基因的调控机制。仅在SCM-1生产T细胞系中，通过环孢菌素A敏感机制，仅在SCM-1生产T细胞系中用抗CD3或PHA加PMA激活后，即可强烈诱导受0.7 kb 5'侧翼区域控制的荧光素酶报道基因。发现相对于主要转录起始位点位于-108至-95nt的称为E1的元件对于启动子活性至关重要。在使用E1寡核苷酸作为探针的电泳迁移率变动分析中，未刺激的T细胞和B细胞系的核提取物形成了称为复合物I的组成性复合物，而受刺激的SCM-1生产者T细胞系的核提取物形成了称为复合物II的较高迁移率的复合物。在激活后仅在SCM-1生产者T细胞系中看到的从复合物I到复合物II的转变被环孢菌素A完全抑制。这两种复合物都严重依赖于NF-AT核心序列TTTCC。 E1元素，并被抗NF-ATp部分超移。酵母分离的NF-ATp作为E1结合蛋白的单杂交试验，以及将NF-ATp转染到T和B细胞系中，可大大增强激活依赖性SCM-1启动子的活性。总的来说，涉及NF-ATp的独特机制似乎可以调节SCM-1基因的细胞类型特异性和激活依赖性表达。

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