Concerted regulation of the porcine steroidogenic acute regulatory protein gene promoter activity by follicle-stimulating hormone and insulin-like growth factor I in granulosa cells involves GATA-4 and CCAAT/enhancer binding protein beta

Endocrinology. 2004 Jul;145(7):3122-34. doi: 10.1210/en.2003-1719. Epub 2004 Apr 1.


We previously demonstrated that FSH alone or in combination with IGF-I activated the porcine steroidogenic acute regulatory protein gene promoter in a concerted manner in primary cultures of granulosa cells. Studies were undertaken to further delineate cis- and trans-acting elements of the porcine promoter and mechanisms mediating FSH stimulation and its augmentation by IGF-I. Mutation of several putative regulatory elements localized hormone-stimulated activity to the highly conserved GATA site and identified novel nucleotides downstream as a functional CCAAT/enhancer binding protein (C/EBP)beta site. In granulosa cell nuclear extracts, GATA-4 and C/EBPbeta formed a high-molecular-weight complex with an oligonucleotide spanning -76 to -32 bp of the porcine promoter. The intensity of this high-molecular-weight complex was increased in granulosa cell nuclear extracts by treatment with FSH alone and was enhanced with the combination of FSH and IGF-I at 2-3 h of treatment. GATA-4 and C/EBPbeta proteins were uniformly expressed with all treatments at time points associated with increased DNA binding. Treatment (2 h) with FSH alone or FSH + IGF-I increased phosphorylation of GATA-4 on a protein kinase A consensus site. The 38-kDa isoform of C/EBPbeta exhibited greater phosphorylation with FSH + IGF-I treatment. Porcine luteal cell nuclear extracts also demonstrated GATA-4 and C/EBPbeta binding to the -76 to -32 bp region of the promoter providing evidence for their cooperation in vivo. These data indicate that GATA-4 and C/EBPbeta are both required for FSH +/- IGF-I stimulation of the porcine steroidogenic acute regulatory protein gene promoter in homologous granulosa cell cultures.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • CCAAT-Enhancer-Binding Protein-beta / metabolism*
  • Cells, Cultured
  • DNA-Binding Proteins / metabolism*
  • Female
  • Follicle Stimulating Hormone / pharmacology*
  • GATA4 Transcription Factor
  • Granulosa Cells / cytology
  • Granulosa Cells / drug effects
  • Granulosa Cells / physiology*
  • Insulin-Like Growth Factor I / pharmacology*
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Phosphoproteins / genetics*
  • Phosphorylation
  • Promoter Regions, Genetic / physiology
  • Sus scrofa
  • Transcription Factors / metabolism*
  • Transfection


  • CCAAT-Enhancer-Binding Protein-beta
  • DNA-Binding Proteins
  • GATA4 Transcription Factor
  • Phosphoproteins
  • Transcription Factors
  • steroidogenic acute regulatory protein
  • Insulin-Like Growth Factor I
  • Follicle Stimulating Hormone