Fig. 1.

Human glucocorticoid receptor cDNA sequencing strategy and schematic representation of cDNA clones, a, The composite cDNA for the α glucocorticoid receptor is represented at the top, with noncoding (lines) and coding (stippled portion) sequences indicated. Common 6-nucleotide restriction enzyme sites are shown. Overlapping cDNA inserts used to determine the sequence are shown: arrows beneath the regions sequenced show the direction and extent of sequencing. The dashed line at the 3’ end of OB10 indicates divergent sequence. Numbers refer to nucleotide positions in OB10 relative to the 5’-most transcribed sequence, b, cDNAs encoding the α and β forms of the receptor (OB7 and OB10, respectively). The 5’ end of OB7 (broken lines) is contributed by the OB10 clone. Protein-coding information is represented by wide bars; untranslated sequences are indicated by thin bars. Nucleotides and amino acids are numbered above and below the coding sequence, respectively. Common DNA sequences extend to nucleotide 2,313 (amino-acid residue 727), at which point the α- and β-receptor forms diverge, with the α cDNAs (OB12, OB7) continuing in an open reading frame for 150 nucleotides (50 amino acids) and the β cDNA (OB10) continuing for 45 nucleotides (15 amino acids; see Fig. 3). Hexanucleotide signals (AATAAA) just upstream of the poly(A) in the clones are indicated, with the first hexanucleotide in OB7 serving as poly(A) in OB12

Methods. The inserts hGR1.2, hGR2.9 and hGR5.16 were isolated from a λ gt11 IM-9 lymphoid cell cDNA library as described previously⁴². Two clones were isolated from cDNA libraries constructed by H. Okayama in pcD (ref. 44) using poly(A)+ mRNA from GM637 human fibroblasts (OB7) and primary human fibroblasts (OB10). Screening was performed with the hGR1.2 cDNA, radiolabelled by nick-translation with ³²P-dCTP. Sequences were determined by the chemical cleavage method of Maxam and Gilbert⁴⁵.