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- Title
A novel splicing silencer generated by DMD exon 45 deletion junction could explain upstream exon 44 skipping that modifies dystrophinopathy.
- Authors
Dwianingsih, Ery Kus; Malueka, Rusdy Ghazali; Nishida, Atsushi; Itoh, Kyoko; Tomoko Lee; Mariko Yagi; Iijima, Kazumoto; Takeshima, Yasuhiro; Matsuo, Masafumi
- Abstract
Duchenne muscular dystrophy (DMD), a progressive muscle-wasting disease, is mostly caused by exon deletion mutations in the DMD gene. The reading frame rule explains that out-of-frame deletions lead to muscle dystrophin deficiency in DMD. In outliers to this rule, deletion junction sequences have never previously been explored as splicing modulators. In a Japanese case, we identified a single exon 45 deletion in the patient's DMD gene, indicating out-of-frame mutation. However, immunohistochemical examination disclosed weak dystrophin signals in his muscle. Reverse transcription-PCR amplification of DMD exons 42 to 47 revealed a major normally spliced product with exon 45 deletion and an additional in-frame product with deletion of both exons 44 and 45, indicating upstream exon 44 skipping. We considered the latter to underlie the observed dystrophin expression. Remarkably, the junction sequence cloned by PCR walking abolished the splicing enhancer activity of the upstream intron in a chimeric doublesex gene pre-mRNA in vitro splicing. Furthermore, antlsense oligonucleotides directed against the junction site counteracted this effect. These Indicated that the junction sequence was a splicing silencer that induced upstream exon 44 skipping. It was strongly suggested that creation of splicing regulator is a modifier of dystrophinopathy.
- Subjects
RNA splicing; DUCHENNE muscular dystrophy; DELETION mutation; OPEN reading frames (Genetics); EXONS (Genetics); REVERSE transcriptase polymerase chain reaction; GENE amplification
- Publication
Journal of Human Genetics, 2014, Vol 59, Issue 8, p423
- ISSN
1434-5161
- Publication type
Article
- DOI
10.1038/jhg.2014.36