Tissue-specific transcriptional initiation of the CYP19 genes in rainbow trout, with analysis of splicing patterns and promoter sequences

Abstract

The rainbow trout ( Oncorhynchus mykiss Walbaum) genome contains three separate CYP19 genes for distinct isoforms of cytochrome P450arom: CYP19A encoding the prevalently ovarian isoform P450aromA, and CYP19B-I and II, encoding forms I and II of the mainly cerebral variant P450aromB. RNA Ligase-Mediated 5′-Rapid Amplification of cDNA Ends analysis was used to determine the 5′-untranslated terminal regions (5′-UTRs) of the corresponding mRNAs, which are actually all expressed in the ovary, brain and gills. CYP19A is transcribed at different transcription start sites (TSSs) in each tissue, the most distal TSS being found in the brain, the intermediate one in the gills, and the proximal one in the ovary. CYP19B-I also displays tissue-specific TSSs, but transcripts undergo three distinct splicing patterns: the same pattern as previously reported for the brain and occurring also in the gills, and two novel patterns, established in the ovary and brain, which include two cryptic 3′-splice sites in intron 1, leading to the inclusion of intronic sequences of 92/94 and 66 b in the 5′-UTRs. Lastly, the CYP19B-II transcript in the ovary shows the same splicing pattern previously described for the brain. A PCR-based gene walking strategy was used to explore the promoter regions of the rainbow trout CYP19 genes, which were found to contain potential binding sites for a variety of transcription factors.