Transcriptional regulation of dehydroepiandrosterone sulfotransferase (SULT2A1) by estrogen-related receptor α
Transcriptional regulation of dehydroepiandrosterone sulfotransferase (SULT2A1) by estrogen-related receptor α
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DOI：
10.1210/en.2004-1619
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发表时间：
2005-08-01
期刊：
影响因子：
4.8
通讯作者：
Rainey, WE
中科院分区：
文献类型：
--
作者：
Seely, J;Amigh, KS;Rainey, WE
雌激素相关受体（ERR α， - β和- γ）是孤儿核受体（指定为NR3B1， NR3B2和NR3B3）的一个亚家族，在结构和功能上与雌激素受体α和β相关。在这里，我们测试ERR α调节基因的转录编码参与肾上腺类固醇生产的酶的假设。Real-time RT-PCR首次用于测定人体各种组织中ERR α mRNA的水平。成人肾上腺中ERR α转录物的水平与心脏中相似，而心脏是高度表达ERR α的。然后用Western blotting和免疫组织化学方法证实ERR α在成人肾上腺中的表达。为了研究ERR α对甾体生成能力的影响，我们使用了含有甾体生成急性调节蛋白（StAR）、胆固醇侧链切割（CYP11A）、3 β -羟基类固醇脱氢酶II型（HSD3B2）、17 α -羟化酶/17,20-裂解酶（CYP17）和脱氢表雄酮磺转移酶（SULT2A1） 5'侧侧区域的报告基因构建。将这些报告基因结构与野生型ERR α或VP16-ERR α表达载体共转染后，发现ERR α在CYP17和SULT2A1的侧链DNA驱动下增强了报告基因活性。SULT2A1启动子活性对ERR α和VP16-ERR α最敏感，活性分别增加2.6倍和79.5倍。ERR α对SULT2A1的影响大于对甾体生成因子1 （SF1）的刺激。转染SULT2A1基因的5'侧DNA序列缺失和EMSA实验表明，存在三个与SF1结合位点序列相似的功能调节顺式元件。综上所述，ERR α在肾上腺中的表达及其对SULT2A1的调控表明，这种孤儿受体在肾上腺类固醇生成的调控中发挥了重要作用。
The estrogen-related receptors (ERR alpha, -beta, and -gamma) are a subfamily of orphan nuclear receptors ( designated NR3B1, NR3B2, and NR3B3) that are structurally and functionally related to estrogen receptors alpha and beta. Herein we test the hypothesis that ERR alpha regulates transcription of the genes encoding the enzymes involved in adrenal steroid production. Real-time RT-PCR was first used to determine the levels of ERR alpha mRNA in various human tissues. Adult adrenal levels of ERR alpha transcript were similar to that seen in heart, which is known to highly express ERR alpha. Expression of ERR alpha in the adult adrenal was then confirmed using Western blotting and immunohistochemistry. To examine the effects of ERR alpha on steroidogenic capacity we used reporter constructs with the 5'-flanking regions of steroidogenic acute regulatory protein ( StAR), cholesterol side-chain cleavage (CYP11A), 3 beta-hydroxysteroid dehydrogenase type II ( HSD3B2), 17 alpha-hydroxylase/17,20-lyase (CYP17), and dehydroepiandrosterone sulfotransferase (SULT2A1). Cotransfection of these reporter constructs with wild-type ERR alpha or VP16-ERR alpha expression vectors demonstrated ERR alpha enhanced reporter activity driven by flanking DNA from CYP17 and SULT2A1. SULT2A1 promoter activity was most responsive to the ERR alpha and VP16-ERR alpha, increasing activity 2.6- and 79.5-fold, respectively. ERR alpha effects on SULT2A1 were greater than the stimulation seen in response to steroidogenic factor 1 (SF1). Transfection of serial deletions of the 5'-flanking DNA of the SULT2A1 gene and EMSA experiments indicated the presence of three functional regulatory cis-elements which shared sequence similarity to binding sites for SF1. Taken together, the expression of ERR alpha in the adrenal and its regulation of SULT2A1 suggest an important role for this orphan receptor in the regulation of adrenal steroid production.