Quantitative analysis of CAPN3 transcripts in LGMD2A patients: Involvement of nonsense-mediated mRNA decay

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• 作者：Kristý ; na Stehl&#xed ; kov&#xe1 ; Eva Zapletalov&#xe1 ; Jana Sedl&#xe1 ; č ; kov&#xe1 ; Mark&#xe9 ; ta Hermanov&#xe1 ; Petr Vondr&#xe1 ; č ; ek ; Tat’ ; &#xe1 ; na Mař ; &#xed ; kov&#xe1 ; Radim M
• 关键词：LGMD2A ; CAPN3 ; Calpain-3 ; Real-time RT-PCR ; Nonsense-mediated mRNA decay
• 刊名：Neuromuscular Disorders
• 出版年：2007
• 期刊代码：203_09608966
• 类别：med
• 出版时间：February 2007
• 卷：17
• 期：2
• 页码：143-147
• 文件大小：219 K

摘要

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 occurence of a missense mutation or an in-frame deletion at the mRNA level although the DNA was heterozygous for this mutation in conjuction 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.