title: Congenital adrenal hyperplasia: the molecular basis of 21-hydroxylase deficiency in H-2(aw18) mice.
abstract: The mouse strain H-2(aw18) shows typical characteristics of 21-hydroxylase deficiency (21-OHD). A deletion of the active Cyp21a1 gene has been postulated; however, the changes on the nucleotide level are still unknown. To investigate whether this animal model, the only one available, is suitable for studying congenital adrenal hyperplasia in man, a detailed analysis of the Cyp21 locus has been performed to ascertain the genetic cause of 21-OHD in H-2(aw18) mice. We demonstrate that 21-OHD is caused by unequal crossing over between the active Cyp21a1 gene and the pseudogene resulting in a hybrid Cyp21a1-Cyp21a2-p gene including a partial deletion of Cyp21a1. Next to several pseudogene-specific point mutations, various novel missense mutations and a nonsense mutation are present. Enzyme activity for each point mutation has been determined in vitro and the structure-function relationship has been studied by sequence conservation analysis and a three-dimensional murine 21-hydroxylase protein (Cyp21) structure model. The mutations are classified in three classes: I, no or minor decrease in enzyme activity: R238Q, P465L, R361K, A362V, P458L; II, loss of enzyme activity caused by inefficient electron flux: R346H, R400C; III, loss of activity due to deficient substrate binding: I462F, L464F. The combination of in vitro protein expression and three-dimensional structure modeling provides a valuable tool to understand the role of the different mutations and polymorphisms on the resulting enzyme activity. The underlying genetic mechanisms are also known to be responsible for 21-OHD in humans, so rodent 21-OHD turns out to be an excellent genetic model for studying the human disease.
authors.affiliation: Division of Paediatric Endocrinology, Department of Paediatrics, Christian-Albrechts-University of Kiel, Schwanenweg 20, D-24105 Kiel, Germany.
authors.forename: Felix G
authors.initials: FG
authors.lastname: Riepe
authors.forename: Stephan
authors.initials: S
authors.lastname: Tatzel
authors.forename: Wolfgang G
authors.initials: WG
authors.lastname: Sippell
authors.forename: Jürgen
authors.initials: J
authors.lastname: Pleiss
authors.forename: Nils
authors.initials: N
authors.lastname: Krone
mesh_terms: D000312:Adrenal Hyperplasia, Congenital / Q000235:genetics* / Q000503:physiopathology*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D018389:Codon, Nonsense; D003434:Crossing Over, Genetic; D004195:Disease Models, Animal; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D005838:Genotype; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D018895:Microsatellite Repeats; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D020125:Mutation, Missense; D016133:Polymerase Chain Reaction; D017434:Protein Structure, Tertiary; D011544:Pseudogenes / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D013255:Steroid 21-Hydroxylase / Q000737:chemistry* / Q000235:genetics*; D013329:Structure-Activity Relationship
chemical_list: D018389:Codon, Nonsense; D012333:RNA, Messenger; D013255:Steroid 21-Hydroxylase