Transcriptional regulation of the rat prostaglandin endoperoxide synthase 2 gene in granulosa cells. Evidence for the role of a cis-acting C/EBP beta promoter element.

摘要: The promoter of the rat prostaglandin endoperoxide synthase 2 (PGS-2) gene has recently been shown to confer gonadotropic hormone (follicle-stimulating (FSH), luteinizing (LH), cAMP) inducibility when ligated a CAT (chloramphenicol acetyltransferase) reporter and transfected into primary cultures differentiated granulosa cells. To delineate cis-acting elements trans-activating factors mediating this response, deletions active region (-192/-53 base pairs upstream transcriptional start site) were tested for their ability bind protein cell nuclear extracts activate activity. Electrophoretic mobility shift assays revealed that DNA subregion -142/-120 inhibited protein/DNA binding observed between labeled PGS-2 fragment -192/-53. acting element C/EBP beta, 5‘-TTATGCAAT-3‘. Point mutations within beta abolished resulted in 50% loss forskolin/LH/FSH mRNA induced rapidly cells vivo by an ovulatory dose human chorionic gonadotropin (hCG). Collectively, these results indicate C/EPB appears play key role regulating induction prior ovulation.