P450c11B3 mRNA, Transcribed from a Third P450c11 Gene, Is Expressed in a Tissue-specific, Developmentally, and Hormonally Regulated Fashion in the Rodent Adrenal and Encodes a Protein with Both 11-Hydroxylase and 18-Hydroxylase Activities

The rat genome contains four P450c11 genes. One of these (CYP11B1) encodes P450c11 beta, which is the steroid 11 beta-hydroxylase found solely in the adrenal zona fasciculata/reticularis, and is responsible for the conversion of 11-deoxycorticosterone to corticosterone. A second P450c11 gene (CYP11B2) encodes P450c11AS, which is the aldosterone synthase found solely in the adrenal zona glomerulosa. P450c11AS has three activities, 11 beta-hydroxylase, 18-hydroxylase, and 18-oxidase, and is responsible for the conversion of 11-deoxycorticosterone to aldosterone. Recently, two more rat P450c11 genes, P450c11B3 and P450c11B4, were cloned. P450c11B4 appears to be a pseudogene, as two exons are replaced by unrelated DNA. P450c11B3 closely resembles P450c11 beta in mRNA and encoded amino acid sequences, predicting a protein of 498 amino acids. However, the expression of this mRNA and protein have not been demonstrated to date. We now demonstrate that this P450c11B3 mRNA is expressed in the adrenal gland several days after birth and is not expressed during fetal development or in the adult rat adrenal. Like P450c11 beta mRNA, P450c11B3 mRNA is expressed in the zona fasciculata/reticularis and not in the zona glomerulosa. However, the regulation of P450c11B3 mRNA expression is different from that of P450c11 beta mRNA, in that its abundance is decreased by ACTH in a sex-dependent fashion. Transfection of eukaryotic cells with a vector expressing P450c11B3 shows that this form of P450c11 can convert 11-deoxycorticosterone (DOC) to corticosterone and thus has the same enzymatic activity as P450c11 beta. In addition, P450c11B3 can convert DOC to 18-OH DOC and corticosterone to 18-OH corticosterone and thus has 18-hydroxylase activity similar to P450c11AS, but it lacks detectable 18-oxidase activity. Thus, P450c11B3 catalyzes 11 beta- and 18-hydroxylation and thus has a spectrum of activities midway between P450c11 beta and P450c11AS.