TY - JOUR
T1 - Canine COL1A2 mutation resulting in C-terminal truncation of pro-α2(I) and severe osteogenesis imperfecta
AU - Campbell, Bonnie G.
AU - Wootton, Joyce A.M.
AU - Macleod, James N.
AU - Minor, Ronald R.
PY - 2001
Y1 - 2001
N2 - 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.
AB - 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.
KW - COL1A2
KW - Canine
KW - Frameshift
KW - Osteogenesis imperfecta
KW - Pro-α2(I)
UR - http://www.scopus.com/inward/record.url?scp=0034999307&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034999307&partnerID=8YFLogxK
U2 - 10.1359/jbmr.2001.16.6.1147
DO - 10.1359/jbmr.2001.16.6.1147
M3 - Article
C2 - 11393792
AN - SCOPUS:0034999307
SN - 0884-0431
VL - 16
SP - 1147
EP - 1153
JO - Journal of Bone and Mineral Research
JF - Journal of Bone and Mineral Research
IS - 6
ER -