IDENTIFICATION OF A LIVER-SPECIFIC PROMOTER FOR THE OVINE GROWTH-HORMONE RECEPTOR

Growth hormone (GH) receptor cDNA clones from several species are characterized by heterogeneity in the 5' untranslated region (5'UT). This has been attributed to different promoters directing the expression of the gene from exons encoding 5'UT's which are alternatively spliced onto a common splice acceptor 11 basepairs (bp) upstream of the initiating AUG on exon 2. The following study identifies exon 1A of the ovine (o) GH receptor gene, corresponding to the 5'UT of a developmentally regulated, liver-specific transcript. Exon 1A spans 206 bp at a position 17 kilobases (kb) upstream of exon 2. Sequencing of the 669 bp region 5' to the transcription initiation site (+1) reveals a TATA box at - 31, a CCAAT box at - 88, and putative binding sites for several transcription factors involved in liver-specific gene expression. Two repetitive sequence elements are located in the 5' and 3' flanking regions of exon 1A. Functional analysis of the 4.5 kb region upstream of exon 1A was performed by transfecting the human hepatoma cell line HuH7 with luciferase reporter gene constructs. Positive and negative regulatory regions are identified, with basal promoter activity within 473 bp of the transcription initiation site. A 47 bp region containing putative binding sites for the activated glucocorticoid receptor and C/EBP-like proteins, between - 180 and - 133, is essential for transcriptional activation.