caa a22 4500 397575262 CHVBK 20180308164854.0 cr unu---uuuuu 161202e199607 xx s 000 0 eng 10.1093/hmg/5.7.995 doi (NATIONALLICENCE)oxford-10.1093/hmg/5.7.995 A Model of mRNA Splicing in Adult Lysosomal Storage Disease (Glycogenosis Type II) [Elektronische Daten] [Nina Raben, Ralph C. Nichols, Frank Martiniuk, Paul H. Plotz] Glycogenosis type II is a recessively inherited disorder caused by mutations in the acid maltase (GAA) gene. Clinically, three different phenotypes are recognized: infantile, juvenile and adult forms. A majority of compound heterozygous adult-onset patients carry a t-13g mutation in intron 1 associated with splicing out the first coding exon (exon 2). We have studied the mechanism of this mutation in a model system with wild-type and mutant minigenes expressed in a GAA deficient cell line. We have demonstrated that the mutation does not prevent normal splicing; low levels of correctly spliced mRNA are generated with the mutant construct. The data explain why the mutation is restricted to a milder, adult-onset phenotype. We also demonstrate that splicing out of exon 2 occurs with the wild-type construct, and thus represents alternative splicing which takes place in normal cells. Three splice variants (SV1, SV2 and SV3) are made with both the mutant and the wild-type constructs. Furthermore, as shown by RNAse protection assay, these mRNA variants are less abundant with the mutant construct. Thus, a major effect of the mutation appears to be a low splicing efficiency, since the total amount of all the transcripts generated from the mutant construct is reduced compared with the wild type. The removal of ∼90% of the intron 1 (2.6 kb) sequence resulted in a dramatic increase in the levels of correctly spliced mRNA, indicating that the intron may contain a powerful transcriptional repressor. © 1996 Oxford University Press Raben Nina Arthritis and Rheumatism Branch, National Institute of Arthritis, Musculoskeletal, and Skin Diseases, National Institutes of Health, Bethesda, Maryland, USA aut Nichols Ralph C. Arthritis and Rheumatism Branch, National Institute of Arthritis, Musculoskeletal, and Skin Diseases, National Institutes of Health, Bethesda, Maryland, USA aut Martiniuk Frank Department of Medicine Pulmonary Division, New York University Medical Center, New York, USA aut Plotz Paul H. Arthritis and Rheumatism Branch, National Institute of Arthritis, Musculoskeletal, and Skin Diseases, National Institutes of Health, Bethesda, Maryland, USA aut Human Molecular Genetics Oxford University Press 5/7(1996-07), 995-1000 0964-6906 5:7<995 1996 5 hmg https://doi.org/10.1093/hmg/5.7.995 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/hmg/5.7.995 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Raben Nina Arthritis and Rheumatism Branch, National Institute of Arthritis, Musculoskeletal, and Skin Diseases, National Institutes of Health, Bethesda, Maryland, USA aut NATIONALLICENCE 700 1- Nichols Ralph C. Arthritis and Rheumatism Branch, National Institute of Arthritis, Musculoskeletal, and Skin Diseases, National Institutes of Health, Bethesda, Maryland, USA aut NATIONALLICENCE 700 1- Martiniuk Frank Department of Medicine Pulmonary Division, New York University Medical Center, New York, USA aut NATIONALLICENCE 700 1- Plotz Paul H. Arthritis and Rheumatism Branch, National Institute of Arthritis, Musculoskeletal, and Skin Diseases, National Institutes of Health, Bethesda, Maryland, USA aut NATIONALLICENCE 773 0- Human Molecular Genetics Oxford University Press 5/7(1996-07), 995-1000 0964-6906 5:7<995 1996 5 hmg Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford