The promoter/regulatory region of the porcine P-450 cholesterol side chain cleavage (P450scc) gene was cloned from a porcine genomic library. This gene contains a GC-rich region (-130-100) that mediates insulin-like growth factor I (IGF-I) and cAMP stimulation of gene expression in porcine granulosa cells. Stimulation of gene expression by cAMP occurs in 6 h, while IGF-I stimulation occurs in 24-48 h. This region is also responsive to insulin but not fibroblast growth factor. The effects of IGF-I and cAMP on gene expression in porcine granulosa cells are additive. In Y1 adrenal cells, the same region is responsive to cAMP but not to IGF-I. Gel shift assays using an oligonucleotide of this region and nuclear extract protein from porcine granulosa and Y1 adrenal cells identified three DNA-protein complexes (C1- C3). The binding activity of the complexes did not change with IGF-I or forskolin treatment of granulosa cells. Mutational analysis results were consistent with IGF-I regulating gene expression through the C2 DNA protein complex. Moreover, this complex binds differently in gel shift assay to mutant oligonucleotides with porcine and Y1 and nuclear extract protein. We conclude that IGF-I stimulates porcine P450scc gene expression through a distinct, cell-specific protein complex binding to a GC-rich domain.
|Original language||English (US)|
|Number of pages||9|
|Journal||Journal of Biological Chemistry|
|State||Published - Oct 14 1994|
ASJC Scopus subject areas
- Molecular Biology
- Cell Biology