Ad4BP/SF-1 regulates cyclic AMP-induced transcription from the proximal promoter (PII) of the human aromatase P450 (CYP19) gene in the ovary

Aromatase P450, which is responsible for the metabolism of C₁₉ steroids to estrogens, is expressed in the pre-ovulatory follicles and corpora lutea of ovulatory women by means of a promoter proximal to the start of translation (PII). To understand how this transcription is controlled by cAMP, we constructed chimeric constructs containing deletion mutations of the proximal promoter 5'-flanking DNA fused to the rabbit β-globin reporter gene. Assay of reporter gene transcription in transfected bovine granulosa and luteal cells revealed that cAMP-stimulated transcription was lost upon deletion from -278 to -100 base pairs, indicating the presence of a functional cAMP-responsive element in this region; however, no classical cAMP-responsive element was found. Mutation of an AGGTCA motif located at - 130 base pairs revealed that this element is crucial for cAMP-stimulated reporter gene transcription. When a single copy of this element was placed upstream of a heterologous promoter, it could act as a weak cAMP-response element. Supershift electrophoretic mobility shift assay and UV cross- linking established that Ad4BP/SF-1 binds to this hexameric element. Ad4BP/SF-1 mRNA and protein levels and DNA binding activity are increased in forskolin-treated luteal cells. We conclude that cAMP-stimulated transcription of human aromatase P450 in the ovary is due, at least in part, to increased levels and DNA binding activity of Ad4BP/SF-1.