OXYSTEROL REGULATION OF STEROIDOGENIC ACUTE REGULATORY PROTEIN GENE-EXPRESSION - STRUCTURAL SPECIFICITY AND TRANSCRIPTIONAL AND POSTTRANSCRIPTIONAL ACTIONS

Oxysterols exert a major influence over cellular cholesterol homeostas is. We examined the effects of oxysterols on the expression of steroid ogenic acute regulatory protein (StAR), which increases the delivery o f cholesterol to sterol-metabolizing P450s in the mitochondria, 22(R)- hydroxycholesterol (22(R)-OHC), 25-OHC, and 27-OHC each increased ster oidogenic factor-1 (SF-1)-mediated StAR gene transactivation by simila r to 2-fold in CV-1 cells. In contrast, cholesterol, progesterone, and the 27-OHC metabolites, 27-OHC-5 beta-3-one and 7 alpha,27-OHC, had n o effect. Unlike our findings in CV-1 cells, SF-1-dependent StAR promo ter activity was not augmented by 27-OHC in COS-1 cells, Y-1 cells, Ee Wo choriocarcinoma cells, Chinese hamster ovary (CHO) cells, and human granulosa cells. Studies examining the metabolism of 27-OHC indicated that CV-1 cells formed a single polar metabolite, 3 beta-OH-5-cholest enoic acid from radiolabeled 27-OHC, However, this metabolite inhibite d StAR promoter activity in CV-1, COS-1 and CHO cells. Because 7 alpha ,27-OHC was unable to increase SF-1-dependent StAR promoter activity, we examined 27-OHC 7 alpha-hydroxylase in COS-1 and CHO cells. COS-1 c ells contained high 7 alpha-hydroxylase activity, whereas the enzyme w as undetectable in CHO cells. The hypothesis that oxysterols act in CV -1 cells to increase StAR promoter activity by reducing nuclear levels of sterol regulatory element binding protein was tested. This notion was refuted when it was discovered that sterol regulatory element bind ing protein-1a is a potent activator of the StAR promoter in CV-1, COS -1, and human granulosa cells. Human granulosa and theca cells, which express endogenous SF-1, contained more than 5-fold more StAR protein following addition of 27-OHC, whereas StAR mRNA levels remained unchan ged. We conclude that 1) there are cell-specific effects of oxysterols on SF-1-dependent transactivation; 2) the ability to increase transac tivation is limited to certain oxysterols; 3) there are cell-specific pathways of oxysterol metabolism; and 4) oxysterols elevate StAR prote in levels through posttranscriptional actions.