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An exon skipping-associated nonsense mutation in the dystrophin gene uncovers a complex interplay between multiple antagonistic splicing elements.
Hum Mol Genet. 2006 Mar 15; 15(6):999-1013.HM

Abstract

A nonsense mutation c.4250T>A (p.Leu1417X) in the dystrophin gene of a patient with an intermediate phenotype of muscular dystrophy induces partial in-frame skipping of exon 31. On the basis of UV cross-linking assays and pull-down analysis, we present evidence that the skipping of this exon is because of the creation of an exonic splicing silencer, which acts as a highly specific binding site (UAGACA) for a known repressor protein, hnRNP A1. Recombinant hnRNP A1 represses exon inclusion both in vitro and in vivo upon transient transfection of C2C12 cells with Duchenne muscular dystrophy (DMD) minigenes carrying the c.4250T>A mutation. Furthermore, we identified a downstream splicing enhancer in the central region of exon 31. This region functions as a Tra2beta-dependent exonic splicing enhancer (ESE) in vitro when inserted into a heterologous splicing reporter, and deletion of the ESE showed that incorporation of exon 31 depends on the Tra2beta-dependent enhancer both in the wild-type and mutant context. We conclude that dystrophin exon 31 contains juxtaposed sequence motifs that collaborate to regulate exon usage. This is the first elucidation of the molecular mechanism leading to exon skipping in the dystrophin gene and allowing the occurrence of a milder phenotype than the expected DMD phenotype. The knowledge of which cis-acting sequence within an exon is important for its definition will be essential for the alternative gene therapy approaches based on modulation of splicing to bypass DMD-causing mutations in the endogenous dystrophin gene.

Authors+Show Affiliations

Laboratoire de Génétique Moléculaire, Institut Universitaire de Recherche Clinique (IURC), CHU Montpellier F34000, France.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

16461336

Citation

Disset, A, et al. "An Exon Skipping-associated Nonsense Mutation in the Dystrophin Gene Uncovers a Complex Interplay Between Multiple Antagonistic Splicing Elements." Human Molecular Genetics, vol. 15, no. 6, 2006, pp. 999-1013.
Disset A, Bourgeois CF, Benmalek N, et al. An exon skipping-associated nonsense mutation in the dystrophin gene uncovers a complex interplay between multiple antagonistic splicing elements. Hum Mol Genet. 2006;15(6):999-1013.
Disset, A., Bourgeois, C. F., 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. Human Molecular Genetics, 15(6), 999-1013.
Disset A, et al. An Exon Skipping-associated Nonsense Mutation in the Dystrophin Gene Uncovers a Complex Interplay Between Multiple Antagonistic Splicing Elements. Hum Mol Genet. 2006 Mar 15;15(6):999-1013. PubMed PMID: 16461336.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - An exon skipping-associated nonsense mutation in the dystrophin gene uncovers a complex interplay between multiple antagonistic splicing elements. AU - Disset,A, AU - Bourgeois,C F, AU - Benmalek,N, AU - Claustres,M, AU - Stevenin,J, AU - Tuffery-Giraud,Sylvie, Y1 - 2006/02/06/ PY - 2006/2/8/pubmed PY - 2007/11/6/medline PY - 2006/2/8/entrez SP - 999 EP - 1013 JF - Human molecular genetics JO - Hum Mol Genet VL - 15 IS - 6 N2 - A nonsense mutation c.4250T>A (p.Leu1417X) in the dystrophin gene of a patient with an intermediate phenotype of muscular dystrophy induces partial in-frame skipping of exon 31. On the basis of UV cross-linking assays and pull-down analysis, we present evidence that the skipping of this exon is because of the creation of an exonic splicing silencer, which acts as a highly specific binding site (UAGACA) for a known repressor protein, hnRNP A1. Recombinant hnRNP A1 represses exon inclusion both in vitro and in vivo upon transient transfection of C2C12 cells with Duchenne muscular dystrophy (DMD) minigenes carrying the c.4250T>A mutation. Furthermore, we identified a downstream splicing enhancer in the central region of exon 31. This region functions as a Tra2beta-dependent exonic splicing enhancer (ESE) in vitro when inserted into a heterologous splicing reporter, and deletion of the ESE showed that incorporation of exon 31 depends on the Tra2beta-dependent enhancer both in the wild-type and mutant context. We conclude that dystrophin exon 31 contains juxtaposed sequence motifs that collaborate to regulate exon usage. This is the first elucidation of the molecular mechanism leading to exon skipping in the dystrophin gene and allowing the occurrence of a milder phenotype than the expected DMD phenotype. The knowledge of which cis-acting sequence within an exon is important for its definition will be essential for the alternative gene therapy approaches based on modulation of splicing to bypass DMD-causing mutations in the endogenous dystrophin gene. SN - 0964-6906 UR - https://www.unboundmedicine.com/medline/citation/16461336/An_exon_skipping_associated_nonsense_mutation_in_the_dystrophin_gene_uncovers_a_complex_interplay_between_multiple_antagonistic_splicing_elements_ L2 - https://academic.oup.com/hmg/article-lookup/doi/10.1093/hmg/ddl015 DB - PRIME DP - Unbound Medicine ER -