Intronic hormone response elements mediate regulation of FKBP5 by progestins and glucocorticoids
Intronic hormone response elements mediate regulation of FKBP5 by progestins and glucocorticoids
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DOI：
10.1379/csc-32r.1
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发表时间：
2004-09-01
影响因子：
3.8
通讯作者：
Scammell, JG
中科院分区：
文献类型：
--
作者：
Hubler, TR;Scammell, JG
FKBP51是一种大分子亲免疫蛋白，糖皮质激素、黄体酮和雄激素能强烈增强FKBP51的表达。然而，FKBP51基因（FKBP5）启动子的3.4 kb片段的活性仅被黄体酮微弱地增加，我们在这里表明它对糖皮质激素和雄激素没有反应。使用matinspector对整个FKBP5进行一致激素反应元件（HREs）扫描。我们发现，在大鼠和小鼠FKBP5中保守的2个内含子E区域含有高匹配分数的hre样序列。含有这些区域的脱氧核糖核酸片段（长度约为1kb）被扩增，并在报告基因检测中检测类固醇反应性。FKBP5的内含子E （pIE2）的一个区域对2个异源报告基因具有糖皮质激素和黄体酮的响应性，而另一个不太保守的内含子E （pIE1）区域仅对黄体酮有响应。在pIE2 （pIE1IE2）上游包含pIE1可增强黄体酮反应，但不能增强糖皮质激素反应。含内含子序列的构建体对雄激素均无反应。在pIE1和pIE2中假定的HREs突变消除了激素反应性。电泳迁移位移分析表明，孕激素受体（PR）在pIE1中与HRE结合，而PR和糖皮质激素受体在pIE2中与HRE相互作用。这些数据表明，远端内含子元件显著参与糖皮质激素和孕激素对FKBP5的转录调控。
Expression of FKBP51, a large molecular weight immunophilin, is strongly enhanced by glucocorticoids, progestins, and androgens. However, the activity of a 3.4-kb fragment of the FKBP51 gene (FKBP5) promoter was only weakly increased by progestin and we show here that it is unresponsive to glucocorticoids and androgens. The entire FKBP5 was scanned for consensus hormone response elements (HREs) using Matlnspector. We found that 2 regions of intron E, which are conserved in rat and mouse FKBP5, contain HRE-like sequences with high match scores. Deoxyribonucleic acid fragments (approximately 1 kb in length) containing these regions were amplified and tested in reporter gene assays for steroid responsiveness. One region of intron E of FKBP5 (pIE2) conferred both glucocorticoid and progestin responsiveness to 2 heterologous reporter genes, whereas the other, less-conserved region of intron E (pIE1) was responsive only to progestins. The inclusion of pIE1 upstream of pIE2 (pIE1IE2) enhanced progestin but not glucocorticoid responsiveness. None of the constructs containing intronic sequences was responsive to androgens. Mutation of the putative HREs within pIE1 and pIE2 eliminated hormone responsiveness. Electrophoretic mobility shift assays demonstrated that progesterone receptors (PR) bound to the HRE in pIE1, whereas both PR and glucocorticoid receptors interacted with the HRE in pIE2. These data suggest that distal intronic elements significantly contribute to transcriptional regulation of FKBP5 by glucocorticoids and progestins.