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Proc Natl Acad Sci U S A. 2004 Nov 2;101(44):15603-8. Epub 2004 Oct 22.

Chromatin immunoprecipitation (ChIP) scanning identifies primary glucocorticoid receptor target genes.

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  • 1Departments of Cellular and Molecular Pharmacology and Medicine, University of California-San Francisco, S572D, Genentech Hall, 600 16th Street, San Francisco, CA 94107-2280, USA.

Abstract

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.

PMID:
15501915
[PubMed - indexed for MEDLINE]
PMCID:
PMC524211
Free PMC Article

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