[No authors listed]
Glucocorticoid receptor (GR) is transcribed in a tissue- and cell-specific manner with multiple exon 1 mRNA variants driven by selective promoters. We recently cloned and characterized the 5.3kb proximal promoter sequence of porcine GR gene containing 7 untranslated alternative first exons each processed by a distinct promoter. In this study, we showed tissue-specific expression of total GR and its exon 1 mRNA variants in hippocampus, muscle and liver of pigs. Total GR mRNA was most abundant in liver, followed by muscle and hippocampus in descending order. Among all the GR exon 1 mRNA variants detected, GR exon 1-9/10 and 1-4 were the most predominant variants in all the three tissues. The abundance of GR exon 1-4 mRNA was similar to that of 1-10 in muscle, but was significantly lower than 1-10 in liver and hippocampus. The activities of truncated short (S) and long (L) promoters of respective GR exon 1 mRNA variants were analyzed by luciferase reporter assay in 3 representative cell lines, SY5Y, C2C12 and HepG2. S1-10 and S1-4 demonstrated significantly higher activities than other short promoters in all the cell lines examined. Nevertheless, the strongest activity and cell specificity were detected for L1-10 promoter, which was consistent with the predominant exon 1-9/10 expression in porcine tissues. Moreover, with 3 potential nGRE binding sites, L1-10 promoter was more sensitive to dexamethasone (DEX) in HepG2. Our data provide basic knowledge of the transcriptional mechanism underlying the tissue- and cell-specific expression of porcine GR under basal or ligand-stimulated conditions.
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