The Human Glucocorticoid Receptor as an RNA-Binding Protein: Global Analysis of Glucocorticoid Receptor-Associated Transcripts and Identification of a Target RNA Motif

摘要: Posttranscriptional regulation is emerging as a key factor in glucocorticoid (GC)-mediated gene regulation. We investigated the role of human GC receptor (GR) an RNA-binding protein and its effect on mRNA turnover airway epithelial cells. Cell treatment with potent budesonide accelerated decay CCL2 (t1/2 = 8 ± 1 min versus 62 17 DMSO-treated cells) CCL7 15 4 114 37 min), but not that CCL5 (t1/2=231 266 5 min) BEAS-2B cell line. This was inhibited by preincubation anti-GR Ab, indicating GR itself plays these transcripts. Coimmunoprecipitation biotin pulldown experiments showed associates mRNAs, mRNA. These methods confirmed targeting primary Association localized to 5′ untranslated region further mapped nt 44–60. The collection transcripts associated GR, identified immunoprecipitation GR-mRNA complexes followed microarray analysis, revealed 479 GR. Computational analysis sequence secondary structures yielded GC-rich motif, which shown bind vitro. motif used predict binding additional 7889 results indicate cytoplasmic interacts subset through specific sequences can regulate rates, suggesting novel posttranscriptional for protein.