title: Molecular analysis of the third component of canine complement (C3) and identification of the mutation responsible for hereditary canine C3 deficiency.
abstract: Genetically determined deficiency of the third component of complement (C3) in the dog is characterized by a predisposition to recurrent bacterial infections and to type 1 membranoproliferative glomerulonephritis. The current studies were undertaken to characterize the cDNA for wild-type canine C3 and identify the molecular basis for hereditary canine C3 deficiency. Amplification, cloning, and sequence analysis indicated that canine C3 is highly conserved in comparison with human, mouse, and guinea pig C3. Southern blot analysis failed to show any gross deletions or rearrangements of DNA from C3-deficient animals. Northern blot analysis indicated that the livers of these animals contain markedly reduced quantities of a normal length C3 mRNA. The full-length 5.1-kb canine C3 cDNA was amplified in overlapping PCR fragments. Sequence analysis of these fragments has shown a deletion of a cytosine at position 2136 (codon 712), leading to a frameshift that generates a stop codon 11 amino acids downstream. The deletion has been confirmed in genomic DNA, and its inheritance has been demonstrated by allele-specific oligonucleotide hybridization.
authors.affiliation: Department of Pediatrics, The Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
authors.forename: R
authors.initials: R
authors.lastname: Ameratunga
authors.forename: J A
authors.initials: JA
authors.lastname: Winkelstein
authors.forename: L
authors.initials: L
authors.lastname: Brody
authors.forename: M
authors.initials: M
authors.lastname: Binns
authors.forename: L C
authors.initials: LC
authors.lastname: Cork
authors.forename: P
authors.initials: P
authors.lastname: Colombani
authors.forename: D
authors.initials: D
authors.lastname: Valle
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D003176:Complement C3 / Q000172:deficiency* / Q000235:genetics*; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000737:chemistry; D004283:Dog Diseases / Q000235:genetics*; D004285:Dogs; D006168:Guinea Pigs; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction
chemical_list: D003176:Complement C3; D018076:DNA, Complementary