Transcriptional regulation of human dual specificity protein phosphatase 1 (DUSP1) gene by glucocorticoids.
Transcriptional regulation of human dual specificity protein phosphatase 1 (DUSP1) gene by glucocorticoids.
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DOI：
10.1371/journal.pone.0013754
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发表时间：
2010-10-29
期刊：
影响因子：
3.7
通讯作者：
Wang JC
中科院分区：
文献类型：
--
作者：
Shipp LE;Lee JV;Yu CY;Pufall M;Zhang P;Scott DK;Wang JC
糖皮质激素是有效的抗炎剂，通常用于治疗炎症性疾病。它们通过细胞内糖皮质激素受体（GR）传递信号，该受体在与配体结合后与基因组糖皮质激素反应元件（GRE）结合以调节相关基因的转录。糖皮质激素抑制炎症的一种机制是通过诱导双特异性磷酸酶-1（DUSP1，又名丝裂原激活蛋白激酶磷酸酶-1，MKP-1）基因。我们发现糖皮质激素在 10 分钟内迅速增加 A549 人肺腺癌细胞中 DUSP1 的转录。使用染色质免疫沉淀 (ChIP) 扫描，我们在 DUSP1 转录起始位点上游的 -1421 和 -1118 之间定位了 GR 结合区域。该区域在报告系统中很活跃，诱变分析确定了位于-1337 和-1323 之间的功能性GRE。我们发现糖皮质激素增加了 DNase I 超敏性，降低了核小体密度，并增加了 GRE 周围基因组区域内的组蛋白 H3 和 H4 乙酰化。 ChIP 实验表明 p300 被招募到 DUSP1 GRE，RNA 干扰实验表明 p300 的减少降低了糖皮质激素刺激的 DUSP1 基因表达和组蛋白 H3 过度乙酰化。此外，p300 的过度表达增强了含有 DUSP1 GRE 的报告基因的糖皮质激素刺激活性，而当组蛋白乙酰转移酶结构域突变时，这种共激活效应会受到损害。使用 GR 和 p300 抗体进行的 ChIP-reChIP 实验显示，糖皮质激素处理后 DUSP1 GRE 显着富集，表明 GR 和 p300 处于招募到 DUSP1 GRE 的同一蛋白质复合物中。我们的研究确定了 DUSP1 基因的功能性 GRE。此外，糖皮质激素对 DUSP1 的转录激活需要 p300 和 GRE 周围染色质结构的快速修饰。总体而言，了解糖皮质激素诱导的 DUSP1 基因转录机制可以为炎症性疾病的治疗方法提供见解。
Glucocorticoids are potent anti-inflammatory agents commonly used to treat inflammatory diseases. They convey signals through the intracellular glucocorticoid receptor (GR), which upon binding to ligands, associates with genomic glucocorticoid response elements (GREs) to regulate transcription of associated genes. One mechanism by which glucocorticoids inhibit inflammation is through induction of the dual specificity phosphatase-1 (DUSP1, a.k.a. mitogen-activated protein kinase phosphatase-1, MKP-1) gene. We found that glucocorticoids rapidly increased transcription of DUSP1 within 10 minutes in A549 human lung adenocarcinoma cells. Using chromatin immunoprecipitation (ChIP) scanning, we located a GR binding region between −1421 and −1118 upstream of the DUSP1 transcription start site. This region is active in a reporter system, and mutagenesis analyses identified a functional GRE located between −1337 and −1323. We found that glucocorticoids increased DNase I hypersensitivity, reduced nucleosome density, and increased histone H3 and H4 acetylation within genomic regions surrounding the GRE. ChIP experiments showed that p300 was recruited to the DUSP1 GRE, and RNA interference experiments demonstrated that reduction of p300 decreased glucocorticoid-stimulated DUSP1 gene expression and histone H3 hyperacetylation. Furthermore, overexpression of p300 potentiated glucocorticoid-stimulated activity of a reporter gene containing the DUSP1 GRE, and this coactivation effect was compromised when the histone acetyltransferase domain was mutated. ChIP-reChIP experiments using GR followed by p300 antibodies showed significant enrichment of the DUSP1 GRE upon glucocorticoid treatment, suggesting that GR and p300 are in the same protein complex recruited to the DUSP1 GRE. Our studies identified a functional GRE for the DUSP1 gene. Moreover, the transcriptional activation of DUSP1 by glucocorticoids requires p300 and a rapid modification of the chromatin structure surrounding the GRE. Overall, understanding the mechanism of glucocorticoid-induced DUSP1 gene transcription could provide insights into therapeutic approaches against inflammatory diseases.
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DOI：
10.1016/j.bbapap.2005.08.017
发表时间：
2005-12-30
影响因子：
3.2
作者：
Kaminska, B
通讯作者：
Kaminska, B
影响因子：
20.3
作者：
Bhattacharyya, Sandip;Brown, Diane E.;Muglia, Louis J.
通讯作者：
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影响因子：
16
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影响因子：
5.3
作者：
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GRANNER, DK
影响因子：
10.5
作者：
Bolton, Eric C.;So, Alex Y.;Yamamoto, Keith R.
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Yamamoto, Keith R.