Role of Phosphorylation, Gene Dosage and Dax-1 in SF-1 Mediated Steroidogenesis

Abstract

The mechanisms by which SF-1 (Steroidogenic Factor-1) and Dax-1 (Dosage-sensitive sex reversal-Adrenal hypoplasia congenita critical region on the X chromosome) dictate adrenal-specific transcriptional programs are the focus of this laboratory. SF-1-mediated transcription is upregulated by phosphorylation of serine 203 located in the hinge region of SF-1. An SF-1S203A mutant attenuates SF-1 activation, while substitution of S203 with a charged aspartate (SF-1S203D) results in a dose dependent increase in SF-1 mediated transcription. Ser203 serves as a substrate for Erk2 in vitro and is critical for activation of SF-1 by multiple components of the MAPK pathway. Isoelectric focusing demonstrates multiple immuno-reactive SF-1 species in mouse adrenal and NCI-H295A cell extracts. We propose that differential phosphorylation of SF-1 by various mitogens serves to couple extracellular signals to adrenal-specific transcriptional programs. Mouse studies utilizing SF-1 heterozygous mice explore the in vivo role of SF-1 levels, SF-1 phosphorylation and SF-1 interaction with Dax-1 in adrenal steroidogenesis. SF-1 heterozygous mice exhibit a marked decrease in baseline and post-stress corticosterone with a concomitant increase in ACTH. The role of Dax-1 in these SF-1 dependent processes is explored in compound SF-1 (+/-)/Dax-1 KO mice that exhibit an increase in basal corticosterone and a decrease in basal ACTH compared to simple SF-1 (+/-) mice. These finding are consistent with an inhibitory role for Dax-1 in SF-1 mediated transcription. Mice that express epitope tagged SF-1 (wild type, SF-1S203A and SF-1S203D) are being used to rescue the heterozygous adrenal phenotype and to determine the in vivo role of SF-1 phosphorylation in adrenal function.