Involvement of activator protein-1 in transcriptional regulation of the human μ-opioid receptor gene
Involvement of activator protein-1 in transcriptional regulation of the human μ-opioid receptor gene
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DOI：
10.1124/mol.61.4.800
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发表时间：
2002-04-01
影响因子：
3.6
通讯作者：
Kraus, J
中科院分区：
文献类型：
--
作者：
Börner, C;Höllt, V;Kraus, J
μ-阿片样物质受体介导阿片样物质作用如镇痛、欣快和免疫调节。μ-阿片受体的基因表达可以通过多种物质调节，包括细胞因子、激素和药物。其中一些刺激（E。例如，在一个实施例中，IL-1 β和可卡因）已被证明可激活AP-1转录因子家族的成员。此外，μ-阿片受体基因的转录由佛波酯12-O-十四烷酰基佛波醇-13-乙酸酯（TPA）诱导，佛波酯12-O-十四烷酰基佛波醇-13-乙酸酯（TPA）是蛋白激酶C的激活剂，蛋白激酶C又是AP-1转录因子的激活剂。这表明涉及蛋白激酶C和激活蛋白1（AP-1）转录因子的信号传导途径对于μ-阿片受体基因的特异性表达模式是重要的。在这份报告中，我们表明，TPA激活AP-1以及转录因子核因子κ B（NF κ B）在μ阿片受体表达神经母细胞瘤细胞系SH SY 5 Y。在这些细胞中进行的转染实验中，两种因子都反式激活含有人μ-阿片受体基因启动子的报告基因构建体的表达。通过用特异性NF κ B抑制剂柳氮磺胺吡啶排除TPA对NF κ B的影响，定位AP-1调节元件。两个AP-1元件，其与经典AP-1结合位点各有一个核苷酸不同，被描绘到启动子的位置-2388和-1434。独立于它们的方向，这些元素赋予TPA对异源胸苷激酶启动子的响应性。AP-1结合到这些元素被证实使用电泳迁移率变化和免疫偏移测定。
mu-Opioid receptors mediate such opioid effects as analgesia, euphoria, and immunomodulation. Gene expression of mu-opioid receptors can be modulated by various substances, including cytokines, hormones, and drugs. Some of these stimuli (e. g., IL-1beta and cocaine) have been shown to activate members of the AP-1 transcription factor family. In addition, transcription of the mu-opioid receptor gene is induced by the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA), an activator of protein kinase C, which in turn is an activator of AP-1 transcription factors. This indicates that signaling pathways involving protein kinase C and activator protein 1 (AP-1) transcription factors are important for the specific expression pattern of the mu-opioid receptor gene. In this report, we show that TPA activates AP-1 as well as the transcription factor nuclear factor kappaB (NFkappaB) in the mu-opioid receptor expressing neuroblastoma cell line SH SY5Y. In transfection experiments performed in these cells, both factors trans-activate expression of reporter gene constructs containing the human mu-opioid receptor gene promoter. By excluding the effects of TPA on NFkappaB with the specific NFkappaB inhibitor sulfasalazine, AP-1 regulatory elements were localized. Two AP-1 elements, which differ in one nucleotide each from the classic AP-1 binding site, were delineated to positions -2388 and -1434 of the promoter. Independent of their orientation, these elements conferred TPA responsiveness on the heterologous thymidine kinase promoter. AP-1 binding to these elements was confirmed using electrophoretic mobility shift and immunoshift assays.