Canine COL1A2 mutation resulting in C-terminal truncation of pro-alpha2(I) and severe osteogenesis imperfecta

B G Campbell; J A Wootton; J N Macleod; R R Minor

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Canine COL1A2 mutation resulting in C-terminal truncation of pro-alpha2(I) and severe osteogenesis imperfecta

Journal article

Peer reviewed

Canine COL1A2 mutation resulting in C-terminal truncation of pro-alpha2(I) and severe osteogenesis imperfecta

B G Campbell, J A Wootton, J N Macleod and R R Minor

Journal of bone and mineral research, Vol.16(6), pp.1147-1153

06/2001

Handle:

https://hdl.handle.net/2376/101797

PMID: 11393792

Abstract

Amino Acid Sequence

Protein Subunits

Frameshift Mutation

Hydroxylation

Cells, Cultured

DNA, Complementary - genetics

Molecular Sequence Data

Osteogenesis Imperfecta - genetics

Collagen Type I

Procollagen - metabolism

Molecular Diagnostic Techniques

Animals

Dogs

Fibroblasts

Female

Collagen - genetics

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-alpha2(I) suggested comigration with the similarly sized pro-alpha2(I) derived from the mutant allele. Furthermore, a-chains were overhydroxylated and the ratio of alpha1(I):alpha2(I) was 3.2:1, consistent with the presence of alpha1(I) homotrimers. Analyses of COL1A2 and type I collagen were both consistent with the described heterozygous mutation affecting the pro-alpha2(I) C-propeptide and confirmed a diagnosis of OI.

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Title: Canine COL1A2 mutation resulting in C-terminal truncation of pro-alpha2(I) and severe osteogenesis imperfecta
Creators: B G Campbell - Department of Biomedical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853, USA
J A Wootton
J N Macleod
R R Minor
Publication Details: Journal of bone and mineral research, Vol.16(6), pp.1147-1153
Publisher: United States
Grant note: T32 RR07059-01 / NCRR NIH HHS AR20793 / NIAMS NIH HHS AR08428 / NIAMS NIH HHS
Identifiers: 99900546649601842
Language: English
Resource Type: Journal article

Canine COL1A2 mutation resulting in C-terminal truncation of pro-alpha2(I) and severe osteogenesis imperfecta

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