A(-2)->G transition at the 3' acceptor splice site of IVS17 characterizes the COL2A1 gene mutation in the original Stickler syndrome kindred

作者：Williams CJ^*; Ganguly A; Considine E; McCarron S; Prockop DJ; WalshVockley C; Michels VV

来源：American Journal of Medical Genetics, 1996, 63(3): 461-467.

DOI：10.1002/(SICI)1096-8628(19960614)63:3<461::AID-AJMG9>3.0.CO;2-U

• Stickler syndrome
• COL2A1 mutation
• splice site mutation

Hereditary progressive arthro-ophthalmopathy, or ''Stickler syndrome,'' is an autosomal dominant osteochondrodysplasia characterized by a variety of ocular and skeletal anomalies which frequently lead to retinal detachment and precocious osteoarthritis. A variety of mutations in the COL2A1 gene have been identified in ''Stickler'' families; in most cases studied thus far, the consequence of mutation is the premature generation of a stop codon, We report here the characterization of a COL2A1 gene mutation in the original kindred described by Stickler et al. [1965].
Conformational sensitive gel electrophoresis (CSGE) [Ganguly et al., 1993] was used to screen for mutations in the entire COL2A1 gene in an affected member from the kindred, A prominent heteroduplex species was noted in the polymerase chain reaction (PCR) product from a region of the gene including exons 17 to 20. Direct sequencing of PCR-amplified genomic DNA resulted in the identification of a base substitution at the A(-2) position of the 3' splice acceptor site of IVS17. Sequencing of DNA from affected and unaffected family members confirmed that the mutation segregated with the disease phenotype, Reverse transcriptase-PCR analysis of poly Af RNA demonstrated that the mutant allele utilized a cryptic splice site in exon 18 of the gene, eliminating 16 bp at the start of exon 18, This frameshift eventually results in a premature termination codon. These findings are the first report of a splice site mutation in classical Stickler syndrome and they provide a satisfying historical context in which to view COL2A1 mutations in this dysplasia.

• 出版日期1996-6-14