Chromatin immunoprecipitation (ChIP) scanning identifies primary glucocorticoid receptor target genes
AUTOR(ES)
Wang, Jen-Chywan
FONTE
National Academy of Sciences
RESUMO
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 transciptional 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.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=524211Documentos Relacionados
- Identifying estrogen receptor α target genes using integrated computational genomics and chromatin immunoprecipitation microarray
- RNAPol-ChIP: a novel application of chromatin immunoprecipitation to the analysis of real-time gene transcription
- ChIP Display: novel method for identification of genomic targets of transcription factors
- Hierarchical hidden Markov model with application to joint analysis of ChIP-chip and ChIP-seq data
- Use of Chromatin Immunoprecipitation To Clone Novel E2F Target Promoters