Abstract
Proper, tissue-specific regulation of CYP19, the gene encoding aromatase, the key enzyme of estrogen synthesis, is essential for reproductive processes. Here, we analyzed transcriptional regulation of the porcine CYP19 in female and male gonads and brain by 5’RACE and RT-PCR and comprehensively mapped the pig CYP19 locus by in silico analysis. Our data revealed that the complete locus, including three paralogous copies, CYP19A1, CYP19A2 and CYP19A3, spans approximately 330 kb of the porcine chromosome 1. The locus also harbors the first exon of the Gliomedin gene (GLDN) in reverse orientation. Only transcripts of the CYP19A3 paralog were substantially expressed in gonads and hypothalamus. We identified CYP19A3-associated untranslated exons approximately 160 kb and 50 kb distal from the first codon. The 5´ untranslated regions of transcripts were derived from either a proximal or from one of these distal untranslated exons. Transcripts including only untranslated exons could be amplified from testis, thus suggesting long non-coding transcripts. The data revealed an additional layer of complexity in the regulation of the porcine CYP19 locus. Tissue-specific expression is not only achieved by tissue- and stage-specific expression of the three different CYP19 paralogs, but also by directing the expression of CYP19A3 from different, proximal and distal promoter regions.
Highlights
IntroductionExpression of CYP19, the gene encoding aromatase, the key enzyme of estrogen synthesis, is essential in regulating female as well as male reproductive processes
It is not surprising that tissue-specific transcription of this gene is tightly regulated. This is achieved by the utilization of different, sometimes very distally located, promoter regions, initially recognized in studies that sequenced CYP19 transcripts isolated from different human tissues
The 5 ́RACE products generated from the testis and male hypothalamus samples from 6-week-old boars were sequenced, revealing that the coding sequences (CDS) was clearly derived from
Summary
Expression of CYP19, the gene encoding aromatase, the key enzyme of estrogen synthesis, is essential in regulating female as well as male reproductive processes. Transcripts, proteins and enzyme activity are present in many different male and female tissues, including the ovaries, placenta and testis [1,2,3]. It is not surprising that tissue-specific transcription of this gene is tightly regulated. This is achieved by the utilization of different, sometimes very distally (relative to the first codon) located, promoter regions, initially recognized in studies that sequenced CYP19 transcripts isolated from different human tissues. The use of alternative, tissue-specific promoters results in transcripts with an identical coding sequence comprising nine different exons
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