COMPOUND HETEROZYGOUS MUTATIONS (ARG 239-]STOP, PRO 342-]THR) IN THE CYP17 (P45017-ALPHA) GENE LEAD TO AMBIGUOUS EXTERNAL GENITALIA IN A MALE-PATIENT WITH PARTIAL COMBINED 17-ALPHA-HYDROXYLASE 17,20-LYASE DEFICIENCY

17-alpha-Hydroxylase deficiency is characterized by defects in either or both the 17-alpha-hydroxylase/17,20-lyase activities. We have, for the first time, elucidated the molecular basis of the deficiency in a male pseudohermaphrodite with ambiguous external genitalia resulting from partial combined deficiency of both activities. The patient is found to be a compound heterozygote, carrying two different inherited mutant alleles in the cytochrome P45017-alpha (CYP17) gene. One allele, from his mother, contains a stop codon (TGA) in place of arginine (CGA) at amino acid position 239 in exon 4. Because this occurs at the N-terminal side of the heme binding sequence, the putative resultant truncated protein is nonfunctional. The second allele, from his father, contains a missense mutation encoding the substitution of proline (CCA) by threonine (ACA) at position 342 in exon 6. Reconstruction of this mutation by site-directed mutagenesis into human P45017-alpha cDNA followed by expression in COS 1 cells leads to the same amount of immunodetectable P45017-alpha protein as found with expression of the normal P45017-alpha cDNA, although both the 17-alpha-hydroxylase and 17,20-lyase activities are found to be reduced to 40-45% of those of the normal enzyme. The presence of ambiguous external genitalia in this 46 XY individual indicates that greater than 20% of the total normal 17,20-lyase activity is required for complete virilization in the male.