Abstract
Production of semi-functional dystrophin mRNA from the dystrophin gene encoding a premature stop codon has been shown to modify the severe phenotype of Duchenne muscular dystrophy (DMD). In this study, we report the tissue-specific production of semi-functional dystrophin mRNA via activation of a nonsense mutation-created intraexonic splice acceptor site. In a DMD patient a novel nonsense mutation was identified in exon 42. In his lymphocytes semi-functional dystrophin mRNA with a 63-nucleotide deletion in exon 42 (dys-63) was found to be produced. In vitro splicing assay using hybrid minigenes disclosed that the mutation-created intraexonic splice acceptor site was activated. In his skeletal muscle cells, however, only the authentically spliced dystrophin mRNA was found. This finding identifies the modulation of the splicing of muscle dystrophin mRNA in cases of DMD as a potential target for therapeutic strategies to generate a milder phenotype for this disease.
This is a preview of subscription content, log in via an institution to check access.
References
Adachi K, Takeshima Y, Wada H, Yagi M, Nakamura H, Matsuo M (2003) Heterogous dystrophin mRNAs produced by a novel splice acceptor site mutation in intermediate dystrophinopathy. Pediatr Res 53:125–131
Barbieri AM, Soriani N, Ferlini A, Michelato A, Ferrari M, Carrera P (1996) Seven novel additional small mutations and a new alternative splicing in the human dystrophin gene detected by heteroduplex analysis and restricted RT-PCR heteroduplex analysis of illegitimate transcripts. Eur J Hum Genet 4:183–187
Disset A, Bourgeois CF, Benmalek N, Claustres M, Stevenin J, Tuffery-Giraud S (2006) An exon skipping-associated nonsense mutation in the dystrophin gene uncovers a complex interplay between multiple antagonistic splicing elements. Hum Mol Genet 15:999–1013
Shiga N, Takeshima Y, Sakamoto H, Inoue K, Yokota Y, Yokoyama M, Matsuo M (1997) Disruption of the splicing enhancer sequence within exon 27 of the dystrophin gene by a nonsense mutation induces partial skipping of the exon and is responsible for Becker muscular dystrophy. J Clin Invest 100:2204–2210
Takeshima Y, Yagi M, Wada H, Ishibashi K, Nishiyama A, Kakumoto M, Sakaeda T, Saura R, Okumura K, Matsuo M (2006) Intravenous infusion of an antisense oligonucleotide results in exon skipping in muscle dystrophin mRNA of Duchenne muscular dystrophy. Pediatr Res 59:690–694
Thi Tran HT, Takeshima Y, Surono A, Yagi M, Wada H, Matsuo M (2005) A G-to-A transition at the fifth position of intron 32 of the dystrophin gene inactivates a splice donor site both in vivo and in vitro. Mol Genet Metab 85:213–219
Tran VK, Takeshima Y, Zhang Z, Yagi M, Nishiyama A, Habara Y, Matsuo M (2006) Splicing analysis disclosed a determinant single nucleotide for exon skipping caused by a novel intra-exonic four-nucleotide deletion in the dystrophin gene. J Med Genet (in press)
Acknowledgments
We would like to thank Ms. A. Hosoda for her secretarial help. This work was supported by a grant-in-aid for Scientific Research from the Japan Society for the Promotion of Science; Health, and Labour Sciences Research Grants for Research on Psychiatric and Neurological Diseases and Mental Health; a research grant for Nervous and Mental Disorders from the Ministry of Health, Labour, and Welfare; and the Mitsubishi Foundation.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Tran, V.K., Takeshima, Y., Zhang, Z. et al. A nonsense mutation-created intraexonic splice site is active in the lymphocytes, but not in the skeletal muscle of a DMD patient. Hum Genet 120, 737–742 (2007). https://doi.org/10.1007/s00439-006-0241-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00439-006-0241-y