GLUCOCORTICOID REGULATION OF C-FOS, C-JUN AND TRANSCRIPTION FACTOR AP-1 IN THE ATT-20 CORTICOTROPE CELL

Glucocorticoids (GC) are potent repressors of both basal and corticotropin releasing factor (CRF) stimulated transcription of the proopiomelanocortin (POMC) gene in corticotrope cells of the anterior pituitary. Despite the finding of a novel, high affinity glucocorticoid receptor (GR) binding site within the proximal region of the POMC promoter, the mechanism by which GC inhibit POMC transcription is still uncertain. Recent studies have described mechanisms whereby GC inhibit transcription of other genes via a direct interaction with components of the transcription factor AP-1. Since it has been shown that CRF stimulates c-fos in AtT-20 corticotrope cells, and that c-fos over-expression elevates POMC transcription, the current study has investigated whether GC can repress c-fos and c-jun gene expression and AP-1 DNA binding activity in AtT-20 corticotrope cells. Acute treatment with doses of dexamethasone (DEX) that markedly inhibited nuclear POMC hnRNA had no effect on basal c-fos mRNA expression, but resulted in a transient down regulation of c-jun. In addition, acute DEX pretreatment significantly lowered CRF stimulation of POMC gene expression and attenuated the CRF stimulation of c-fos mRNA by 25%. Although DEX treatment of AtT-20 cells did not affect AP-1 DNA binding capacity of nuclear extracts, DEX pretreatment blunted the stimulation of AP-1 binding in response to CRF. In further studies, nuclear extracts from CRF-treated cells were coincubated with nuclear extracts from control or DEX treated cells. High levels of DEX treated extracts led to a relative repression of CRF-induced AP-1 binding, suggesting that ligand-activated GR may lower available AP-1 levels by direct protein:protein interaction. Finally, the composition of AP-1 in AtT-20 nuclear extracts was found to be heterogeneous, with the variation dependent upon hormonal treatment. These data suggest that in the corticotrope cell relatively high levels of activated GR may influence CRF-induced AP-1 DNA binding via transient genomic actions on basal c-jun and stimulated c-fos and/or via direct protein:protein interactions.