Canine COL1A2 mutation resulting in C-terminal truncation of pro-α2(I) and severe osteogenesis imperfecta

Bonnie G. Campbell, Joyce A.M. Wootton, James N. Macleod, Ronald R. Minor

Research output: Contribution to journalArticlepeer-review

38 Scopus citations

Abstract

RNA and type I collagen were analyzed from cultured skin fibroblasts of a Beagle puppy with fractures consistent with type III osteogenesis imperfecta (OI). In a nonisotopic RNAse cleavage assay (NIRCA), the proband's RNA had a unique cleavage pattern in the region of COL1A2 encoding the C-propeptide. DNA sequence analyses identified a mutation in which nucleotides 3991-3994 ("CTAG") were replaced with "TGTCATTGG." The first seven bases of the inserted sequence were identical to nucleotides 4002-4008 of the normal canine COL1A2 sequence. The resulting frameshift changed 30 amino acids and introduced a premature stop codon. Reverse-transcription polymerase chain reaction (RT-PCR) with primers flanking the mutation site amplified two complementary DNA (cDNA) fragments for the proband and a single product for the control. Restriction enzyme digestions also were consistent with a heterozygous mutation in the proband. Type I procollagen labeled with [3H]proline was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Increased density of pC-α2(I) suggested comigration with the similarly sized pro-α2(I) derived from the mutant allele. Furthermore, α-chains were overhydroxylated and the ratio of α1(I):α2(I) was 3.2:1, consistent with the presence of α1(I) homotrimers. Analyses of COL1A2 and type I collagen were both consistent with the described heterozygous mutation affecting the pro-α2(I) C-propeptide and confirmed a diagnosis of OI.

Original languageEnglish
Pages (from-to)1147-1153
Number of pages7
JournalJournal of Bone and Mineral Research
Volume16
Issue number6
DOIs
StatePublished - 2001

Keywords

  • COL1A2
  • Canine
  • Frameshift
  • Osteogenesis imperfecta
  • Pro-α2(I)

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Orthopedics and Sports Medicine

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