Comprehensive cDNA study and quantitative transcript analysis of mutantOPA1transcripts containing premature termination codons
| Autor: | Bernd Wissinger, Simone Schimpf, Nico Fuhrmann, Simone Schaich |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
endocrine system
DNA Complementary DNA Mutational Analysis Nonsense-mediated decay Mutant Biology medicine.disease_cause GTP Phosphohydrolases Exon Complementary DNA Optic Atrophy Autosomal Dominant Genetics medicine Humans Gene Alleles Genetics (clinical) Mutation Genetic Variation Molecular biology eye diseases Codon Nonsense Allelic heterogeneity Haploinsufficiency |
| Zdroj: | Human Mutation. 29:106-112 |
| ISSN: | 1098-1004 1059-7794 |
| DOI: | 10.1002/humu.20607 |
| Popis: | Autosomal dominant optic atrophy (adOA) is most commonly caused by mutations in the OPA1 gene. There is a considerable allelic heterogeneity among adOA-associated OPA1 mutations, however these mutations have mostly been identified and studied only at the genomic DNA level. Here we report the identification of 22 novel OPA1 mutations and their analysis at the cDNA level along with 15 already known OPA1 mutations. We found that 18 of these mutations cause splice defects that involve either skipping of the adjacent exon or the activation of a cryptic splice site. We also observed a reduced level of the mutant transcript in several adOA subjects. Allele-specific quantification of the transcript steady-state level was performed for 13 different OPA1 mutations applying pyrosequencing to a RT-PCR amplified cSNP (c.2109C>T) in OPA1. Using this new assay we could demonstrate that the majority of OPA1 mutations that lead to a premature termination codon (PTC) undergo nonsense-mediated mRNA decay (NMD). Mutant transcript levels were reduced between 1.25- and 2.5-fold and varied between PTC containing mutations, and between subjects. Our results emphasize the value of cDNA analysis in the characterization of OPA1 mutations and further strengthen the model of haploinsufficiency as a major pathomechanism in OPA1-associated adOA. |
| Databáze: | OpenAIRE |
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