title: Molecular cloning of cDNA and genomic DNA for human 25-hydroxyvitamin D3 1 alpha-hydroxylase.
abstract: The 25-hydroxyvitamin D3 1 alpha-hydroxylase (1 alpha-hydroxylase) is a cytochrome P450 enzyme that catalyzes the conversion of 25-hydroxyvitamin D3 to 1 alpha,25-dihydroxyvitamin D3. This enzyme plays an important role in calcium homeostasis. Here we report the molecular cloning of cDNA and gene for human 1 alpha-hydroxylase. The cDNA clone was obtained from a human kidney cDNA library by cross-hybridization with a previously cloned rat cDNA probe. The cDNA consists of 2469 bp and encodes a protein of 508 amino acids that shows 82.5% sequence identity with the rat enzyme. A computer-aided homology search revealed that 1 alpha-hydroxylase shares a relatively high homology with vitamin D3 25-hydroxylase (about 40% amino acid identity). Northern blot analysis showed that the 2.5-kb mRNA is most abundant in kidney. The gene for human 1 alpha-hydroxylase spans approximately 6 kb, is composed of nine exons, and is present as a single copy. This molecular cloning makes it possible to investigate the genetic mechanism of diseases related to calcium metabolism, including vitamin D-dependency rickets type I.
authors.affiliation: School of Medicine, Keio University, Tokyo, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Monkawa
authors.forename: T
authors.initials: T
authors.lastname: Yoshida
authors.forename: S
authors.initials: S
authors.lastname: Wakino
authors.forename: T
authors.initials: T
authors.lastname: Shinki
authors.forename: H
authors.initials: H
authors.lastname: Anazawa
authors.forename: H F
authors.initials: HF
authors.lastname: Deluca
authors.forename: T
authors.initials: T
authors.lastname: Suda
authors.forename: M
authors.initials: M
authors.lastname: Hayashi
authors.forename: T
authors.initials: T
authors.lastname: Saruta
mesh_terms: D015090:25-Hydroxyvitamin D3 1-alpha-Hydroxylase / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification*; D018076:DNA, Complementary / Q000302:isolation & purification*; D005091:Exons; D005796:Genes; D015698:Genomic Library; D006801:Humans; D007438:Introns; D008297:Male; D008969:Molecular Sequence Data; D009928:Organ Specificity / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger; D004247:DNA; D015090:25-Hydroxyvitamin D3 1-alpha-Hydroxylase