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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Chromatin immunoprecipitation (ChIP) scanning identifies primary glucocorticoid receptor target genes.
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Chromatin immunoprecipitation (ChIP) scanning identifies primary glucocorticoid receptor target genes.

机译：染色质免疫沉淀（ChIP）扫描可识别初级糖皮质激素受体靶基因。

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摘要

The global physiological effects of glucocorticoids are well established, and the framework of transcriptional regulation by the glucocorticoid receptor (GR) has been described. However, the genes directly under GR control that trigger these physiological effects are largely unknown. To address this issue in a single cell type, we identified glucocorticoid-responsive genes in A549 human lung adenocarcinoma cells by microarray analysis and quantitative real-time PCR. Reduction of GR expression by RNA interference diminished the effects of dexamethasone on all tested target genes, thus confirming the essential role of GR in glucocorticoid-regulated gene expression. To identify primary GR target genes, in which GR is a component of the transcriptional regulatory complex, we developed a strategy that uses chromatin immunoprecipitation to scan putative regulatory regions of target genes for sites occupied by specifically bound GR. We screened 11 glucocorticoid-regulated genes, and we identified GR-binding regions for eight of them (five induced and three repressed). Thus, our approach provides a means for rapid identification of primary GR target genes and glucocorticoid-response elements, which will facilitate analyses of transcriptional regulatory mechanisms and determination of hormone-regulated gene networks.
机译：糖皮质激素的整体生理作用已得到充分确立，并且已描述了糖皮质激素受体（GR）进行转录调控的框架。然而，直接受GR控制的触发这些生理效应的基因在很大程度上尚不清楚。为了在单个细胞类型中解决此问题，我们通过微阵列分析和定量实时PCR在A549人肺腺癌细胞中鉴定了糖皮质激素应答基因。通过RNA干扰减少GR表达可减少地塞米松对所有测试靶基因的影响，从而证实GR在糖皮质激素调节基因表达中的重要作用。为了鉴定其中GR是转录调控复合体组成部分的主要GR目标基因，我们开发了一种策略，该策略使用染色质免疫沉淀法扫描靶基因的假定调控区域以寻找特异性结合的GR所占据的位点。我们筛选了11个糖皮质激素调节的基因，我们确定了其中8个的GR结合区域（5个诱导和3个被抑制）。因此，我们的方法为快速鉴定主要GR靶基因和糖皮质激素应答元件提供了一种手段，这将有助于分析转录调控机制和确定激素调控基因网络。

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