Low-stringency hybridization with human glucocorticoid receptor (hGR) complementary DNA was used to isolate a new gene encoding a predicted 107-kilodalton polypeptide. Expression studies demonstrate its ability to bind aldosterone with high affinity and to activate gene transcription in response to aldosterone, thus establishing its identity as the human mineralocorticoid receptor (hMR). This molecule also shows high affinity for glucocorticoids and stimulates a glucocorticoid-responsive promoter. Together the hMR and hGR provide unexpected functional diversity in which hormone-binding properties, target gene interactions, and patterns of tissue-specific expression may be used in a combinatorial fashion to achieve complex physiologic control.

Identification of complementary DNAs encoding the human glucocorticoid receptor predicts two protein forms, of 777 (alpha) and 742 (beta) amino acids, which differ at their carboxy termini. The proteins contain a cysteine/lysine/arginine-rich region which may define the DNA-binding domain. Pure radiolabelled glucocorticoid receptor, synthesized in vitro, is immunoreactive and possesses intrinsic steroid-binding activity characteristic of the native glucocorticoid receptor.

Inhibin, a gonadal protein that preferentially suppresses the secretion of pituitary follicle-stimulating hormone, has been isolated from porcine follicular fluid and characterized as a 32-kDa protein composed of 18-kDa and 14-kDa subunits. In the present work, oligonucleotide probes predicted from amino-terminal inhibin amino acid sequences have been used to isolate, from a porcine ovarian Xgtll cDNA library, clones encoding the 18-kDa subunit, or A chain, of inhibin. DNA sequence analysis showed that the inhibin A chain is initially synthesized as a larger precursor protein and is predicted to be a glycopeptide. Inhibin A-chain mRNA is present specifically in the gonads, and its synthesis can be induced by treatment of the animal with gonadotropins. The porcine probe was used to isolate a human inhibin A-subunit cDNA from a placental cDNA library. The human precursor is highly homologous to its porcine counterpart and is predicted to generate an 18-kDa glycosylated inhibin A subunit.The biological basis for gonadal regulation of pituitary function was formulated in 1923 by Mottram and Cramer (1), who observed hypertrophy of rat pituitary cells following radiation-induced testicular damage. In 1932 McCullaugh (2) demonstrated that the appearance of these hypertrophied cells could be inhibited by the injection of a water-soluble substance derived from bovine testes, and he termed this substance inhibin. Following the discovery of follicle-stimulating hormone (FSH) and luteinizing hormone (LH), the two pituitary hormones known to regulate the development and activity of the gonads, Klinefelter et al. (3) postulated that a testicular factor, inhibin, exerted a specific negative feedback action on pituitary FSH secretion. This hypothesis was substantiated when numerous investigators demonstrated a direct suppression of peripheral FSH in animals treated with steroid-free testicular or ovarian preparations (4-6).

We have isolated and characterized overlapping clones from phage X and cosmid human genomic libraries that predict the entire structure of the gene encoding the precursor to human growth hormone-releasing factor. The gene includes five exons spanning 10 kilobase pairs of human genomic DNA. There appears to be a segregation of distinct functional regions of the GRF precursor and its mRNA into the five exons of the gene. The DNA sequences of all exons, intron/exon boundaries, and 5' and 3' flanking regions are presented. Dot-blot analysis of DNA from high resolution duallaser-sorted human chromosomes indicates that the singlecopy growth hormone-releasing factor gene is located on human chromosome 20.