Heterogeneity of rat type I 5a-reductase cDNA: cloning, expression and regulation by pituitary implants and dihydrotestosterone

摘要

Primer extension analysis reveals the presence of different forms of mRNA species for rat type I 5"-reductase. Using a 5"-reductase cDNA probe to screen the rat liver λgtl 1 cDNA library, we isolated cDNA clones that have 4 additional amino acids in the NH_2-terminal! region as compared with the previously reported .sequence for rat type I 5"-reductase. These four additional amino acids elongate the rat type I 5"-reductase amino acid sequence to 259 amino acids, the same number as in human type I 5α-rreductase with which it shares 60% identity. Expression of the long and short rat type 1 5α-reductase by transfection in human adrenal adenocarcinoma cells, SW-B cells, indicated that the long cDNA encoded a protein with a higher affinity for the substrate than the short cDNA. To determine the effect of pituitary hormones and dihydrotestosterone (DHT), the mRNA levels in the livers of rats treated with pituitary implants, hypophysectomized, castrated, and castrated coupled with DHT treatment were quantified by dot-blot hybridization assay using rat type I 5α-reductase cDNA a.s probes. The results demonstrated that rat type I 5α-reductase mRNA is stimulated by pituitary hormones and castration but is decreased by DHT and hypophysectomy.