IDENTIFICATION OF 3',5'-CYCLIC ADENOSINE-MONOPHOSPHATE RESPONSE ELEMENT AND OTHER CIS-ACTING ELEMENTS IN THE HUMAN ANDROGEN RECEPTOR GENE PROMOTER
IDENTIFICATION OF 3',5'-CYCLIC ADENOSINE-MONOPHOSPHATE RESPONSE ELEMENT AND OTHER CIS-ACTING ELEMENTS IN THE HUMAN ANDROGEN RECEPTOR GENE PROMOTER
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DOI：
10.1210/me.8.1.77
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发表时间：
1994-01-01
影响因子：
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通讯作者：
CHANG, CS
中科院分区：
文献类型：
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作者：
MIZOKAMI, A;YEH, SY;CHANG, CS
雄激素和雄激素受体(AR)在性分化和前列腺增殖中起重要作用。为了研究AR基因的转录调控，我们分离、测序并鉴定了一个2.3kb的AR基因启动子区域。氯霉素乙酰转移酶(CAT)分析和AR基因启动子在人、大鼠和小鼠中的序列同源性搜索揭示了一些潜在的顺式作用元件，包括一个GC盒、一个抑制区和一个富嘌呤元件。用50个碱基对(BP)的双链富嘌呤元件进行缺失分析和凝胶滞留分析表明，这种富嘌呤元件可以与LNCaP和HeLa细胞核提取液中的特定蛋白结合，可能是AR基因转录所必需的。此外，为了研究cAMP对AR基因转录的影响，我们用10 mM(Bu)(2)cAMP处理LNCaP和HeLa细胞。该处理仅诱导了LNCaP细胞CAT活性的几倍，Northern印迹分析和逆转录聚合酶链式反应进一步证实了这种诱导作用。对AR基因启动子的缺失分析表明，在AR基因转录起始点上游530~380bp之间的一个区域，包括一个潜在的cAMP反应元件(Cre)，负责cAMP的诱导。凝胶滞留分析表明，AR/CRE1能与LNCaP细胞核提取液中的特定蛋白结合，与生长抑素/CRE形成不同的结合复合体。
Androgen and androgen receptor (AR) play an important role in sexual differentiation and prostate proliferation. To investigate AR gene transcriptional regulation, a 2.3-kilobase AR gene promoter region was isolated, sequenced, and characterized. Chloramphenicol acetyltransferase (CAT) assay and sequence homology search of AR gene promoter among human, rat, and mouse revealed some potential cis-acting elements, including a GC box, a suppressor region, and a purine-rich element. Deletion analysis and gel retardation assay using a 50-base pair (bp) double-strand purine-rich element showed that this purine-rich element can bind to specific proteins in nuclear extract of LNCaP and HeLa cells and may be essential for AR gene transcription. Furthermore, to investigate the effect of cAMP on AR gene transcription, we treated LNCaP and HeLa cells with 10 mm (Bu)(2)cAMP after transfection with CAT gene reporter plasmids linked to the AR gene promoter. This treatment induced several folds of CAT activity in LNCaP cells only, and the induction was further confirmed at AR mRNA level by Northern blot analysis and reverse transcription-polymerase chain reaction assay. Deletion analysis of the AR gene promoter showed that a region between 530 bp and 380 bp upstream of AR gene transcription initiation site, which includes one potential cAMP response element (CRE), is responsible for cAMP induction. Gel retardation analysis using this CRE (AR/CRE1) showed that AR/CRE1 can bind to specific proteins in nuclear extract of LNCaP cells, which appears to form a different binding complex compared to somatostatin/CRE.