The 25-hydroxyvitamin D 1-alpha-hydroxylase gene maps to the pseudovitamin D deficiency rickets (PDDR) disease locus

Pseudovitamin D-deficiency rickets (PDDR) is an autosomal recessive disorder that may be due to impaired activity of 25-hydroxyvitamin D-1 alpha-hydroxylase, a renal cytochrome P450 enzyme (P450(1 alpha) of the vitamin D pathway. The disease locus for PDDR has been mapped by linkage analysis to 12q13-q14, but the molecular defect underlying the enzyme dysfunction has remained elusive due to the lack of sequence information for the P450(1 alpha) gene (hereafter referred to as 1 alpha-OHase). We have used a probe derived from the rat 25-hydroxyvitamin D-24-hydroxylase (CYP24; 24-OHase) sequence to identify and clone the 1 alpha-OHase cDNA. The full-length 1 alpha-OHase clone of 2.4 kb codes for a protein of predicted Mr 55 kDa, Functional activity of the cloned sequence was assessed using transient transfection, and the production of authentic 1 alpha,25-dihydroxyvitamin D-3 [1 alpha,25(OH)(2)D-3] was confirmed using high performance liquid chromatography fractionation and time-of-night mass spectrometry. The expression of the gene was analyzed in vitamin D-replete animals; treatment with 1 alpha,25(OH)(2)D-3 reduced 1 alpha-OHase transcript levels by 70%, while administration of parathyroid hormone led to a 2-fold increase in the expression of the gene, thus confirming the hormonal regulation previously described using biochemical methods, The rat cDNA was used to obtain a human genomic clone, Interestingly, the human 1 alpha-OHase gene mapped to 12q13.1-q13.3, providing strong evidence that a mutation in the 1 alpha-OHase gene is responsible for the PDDR phenotype, The availability of a cloned sequence for 1 alpha-OHase generates novel tools for the study of the molecular etiology of PDDR, and will allow the investigation of other disturbances of vitamin D metabolism.