P53 STIMULATES PROMOTER ACTIVITY OF THE SGK SERUM GLUCOCORTICOID-INDUCIBLE SERINE/THREONINE PROTEIN-KINASE GENE IN RODENT MAMMARY EPITHELIAL-CELLS/

sgk is a novel member of the serine/threonine protein kinase gene fami ly that is transcriptionally regulated by serum and glucocorticoids in mammary epithelial cells. To functionally determine if the sgk promot er is regulated by the p53 tumor suppressor protein in mammary cells, a series of sgk promoter fragments with 5'-deletions were linked to th e bacterial chloramphenicol acetyltransferase gene (sgk-CAT) and trans iently cotransfected into nontumorigenic NMuMG or transformed Con8Hd6 mammary epithelial cells with p53 expression plasmids. Wild-type p53, but not mutant p53, strongly stimulated sgk promoter activity in both mammary epithelial cell lines. These effects were mediated by specific regions within the sgk promoter containing p53 DNA-binding sites. The sgk p53 sequence at -1380 to -1345 (site IV) was sufficient to confer p53-dependent transactivation to a heterologous promoter, and p53 was capable of binding to this sequence in vitro as assessed by gel shift analysis. In the nontumorigenic NMuMG epithelial cell line, cotransfe ction of wild-type p53 strongly stimulated the activities of both the sgk promoter and the well characterized p53-responsive p21/Waf1 promot er, whereas in Rat a fibroblasts, wild-type p53 repressed the basal ac tivities of both promoters, revealing that sgk and p21/Waf1 are simila rly regulated in a cell type-specific manner. Taken together, these re sults demonstrate that sgk is a new transcriptional target of p53 in m ammary epithelial cells and represent the first example of a hormone-r egulated protein kinase gene with a functionally defined p53 promoter recognition element.