STRUCTURE AND TISSUE-SPECIFIC EXPRESSION OF 3-BETA-HYDROXYSTEROID DEHYDROGENASE/5-ENE-4-ENE ISOMERASE GENES IN HUMAN AND RAT CLASSICAL AND PERIPHERAL STEROIDOGENIC TISSUES

The enzyme 3-beta-hydroxysteroid dehydrogenase/5-ene-4-ene isomerase (3-beta-HSD) catalyzes the oxidation and isomerization of 5-ene-3-beta-hydroxypregnene and 5-ene-hydroxyandrostene steroid precursors into the corresponding 4-ene-ketosteroids necessary for the formation of all classes of steroid hormones. We have recently characterized two types of human 3-beta-HSD cDNA clones and the corresponding genes which encode deduced proteins of 371 and 372 amino acids, respectively, and share 93.5% homology. The human 3-beta-HSD genes containing 4 exons were assigned by in situ hybridization to the p11-p13 region of the short arm of chromosome 1. We have also recently elucidated the structure of three types of rat 3-beta-HSD cDNAs as well as that of one type of 3-beta-HSD from bovine and macaque ovary lambda-gt11 cDNA libraries which all encode 372 amino acid proteins. The human type I 3-beta-HSD is the almost exclusive mRNA species detected in the placenta and skin, while the human type II is the predominant mRNA species in the adrenals, ovaries and testes. The predicted rat type I and type II 3-beta-HSD proteins expressed in adrenals, gonads and adipose tissue share 94% homology while they share 80% similarity with the liver-specific type III 3-beta-HSD. Transient expression of human type I and type II as well as rat type I and type II 3-beta-HSD cDNAs in HeLa human cervical carcinoma cells reveals that 3-beta-ol dehydrogenase and 5-ene-4-ene isomerase activities reside within a single protein and these cDNAs encode functional 3-beta-HSD proteins that are capable of converting 3-beta-hydroxy-5-ene-steroids into 3-keto-4-ene derivatives as well as the interconversion of 3-beta-hydroxy and 3-keto-5-alpha-androstane steroids. We have found that the rat type III mRNA species was below the detection limit in intact female liver while, following hypophysectomy, its accumulation increased to 55% of the levels measured in intact or HYPOX male rats, an increase which can be blocked by administration of ovine prolactin (oPRL). In addition, in female rats, treatment with oPRL for 10 days starting 15 days after HYPOX, markedly decreased ovarian 3-beta-HSD mRNA accumulation accompanied by a similar decrease in 3-beta-HSD activity and protein levels. Treatment with the gonadotropin hCG reversed the potent inhibitory effect of oPRL on these parameters and stimulated 3-beta-HSD mRNA levels in ovarian interstitial cells. In intact females, hCG exerted marked trophic effects on rat corpora lutea with an increase in total ovarian 3-beta-HSD expression and activity. We have also shown that treatment with hCG for 15 days in intact male rats caused a marked increase in testicular 3-beta-HSD expression and activity while glucocorticoids exerted inhibitory effects on these parameters. We have also observed that the ontogeny of 3-beta-HSD expression in human and rat adrenal gland, testis and ovary is closely correlated with steroid hormone biosynthesis, thus suggesting that regulation of the expression of 3-beta-HSD is a limiting step in the biosynthesis of steroids in these tissues.