Quantitative analysis of CAPN3 transcripts in LGMD2A patients: involvement of nonsense-mediated mRNA decay
| Autor: | Jiří Fajkus, Markéta Hermanová, Josef Zamecnik, Petr Vondráček, Tat’ána Maříková, Stanislav Voháňka, Kristýna Stehlíková, Radim Mazanec, Jana Sedláčková, Lenka Fajkusová, Eva Zapletalová |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
Adult
Male Adolescent Genotype Nonsense-mediated decay Nonsense mutation Blotting Western Muscle Proteins Biology medicine.disease_cause Frameshift mutation Denaturing high performance liquid chromatography 03 medical and health sciences 0302 clinical medicine medicine Missense mutation Humans RNA Messenger Child Frameshift Mutation Gene Genetics (clinical) 030304 developmental biology Genetics 0303 health sciences Mutation Calpain Reverse Transcriptase Polymerase Chain Reaction Proteolytic enzymes Infant DNA Molecular biology Introns 3. Good health Neurology Muscular Dystrophies Limb-Girdle Codon Nonsense Child Preschool Pediatrics Perinatology and Child Health Female Neurology (clinical) 030217 neurology & neurosurgery Gene Deletion |
| Zdroj: | Neuromuscular disorders : NMD. 17(2) |
| ISSN: | 0960-8966 |
| Popis: | Limb girdle muscular dystrophy type 2A (LGMD2A) is caused by single or small nucleotide changes widespread along the CAPN3 gene, which encodes the muscle-specific proteolytic enzyme calpain-3. About 356 unique allelic variants of CAPN3 have been identified to date. We performed analysis of the CAPN3 gene in LGMD2A patients at both the mRNA level using reverse transcription-PCR, and at the DNA level using PCR and denaturing high performance liquid chromatography. In four patients, we detected homozygous occurrence of a missense mutation or an in-frame deletion at the mRNA level although the DNA was heterozygous for this mutation in conjunction with a frame-shift mutation. The relationship observed in 12 patients between the quantity of CAPN3 mRNA, determined using real-time PCR, and the genotype leads us to propose that CAPN3 mRNAs which contain frame-shift mutations are degraded by nonsense-mediated mRNA decay. Our results illustrate the importance of DNA analysis for reliable establishment of mutation status, and provide a new insight into the process of mRNA decay in cells of LGMD2A patients. |
| Databáze: | OpenAIRE |
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