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Activation of cryptic splice sites in three patients with chronic granulomatous disease

BACKGROUND: Chronic granulomatous disease (CGD) is a primary immune deficiency caused by mutations in the genes encoding the structural components of the phagocyte NADPH oxidase. As a result, the patients cannot generate sufficient amounts of reactive oxygen species required for killing pathogenic m...

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Autores principales: de Boer, Martin, van Leeuwen, Karin, Hauri‐Hohl, Mathias, Roos, Dirk
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6732321/
https://www.ncbi.nlm.nih.gov/pubmed/31364312
http://dx.doi.org/10.1002/mgg3.854
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author de Boer, Martin
van Leeuwen, Karin
Hauri‐Hohl, Mathias
Roos, Dirk
author_facet de Boer, Martin
van Leeuwen, Karin
Hauri‐Hohl, Mathias
Roos, Dirk
author_sort de Boer, Martin
collection PubMed
description BACKGROUND: Chronic granulomatous disease (CGD) is a primary immune deficiency caused by mutations in the genes encoding the structural components of the phagocyte NADPH oxidase. As a result, the patients cannot generate sufficient amounts of reactive oxygen species required for killing pathogenic microorganisms. METHODS: We analyzed NADPH oxidase activity and component expression in neutrophils, performed genomic DNA and cDNA analysis, and used mRNA splicing prediction tools to evaluate the impact of mutations. RESULTS: In two patients with CGD, we had previously found mutations that cause aberrant pre‐mRNA splicing. In one patient an exonic mutation in a cryptic donor splice site caused the deletion of the 3' part of exon 6 from the mRNA of CYBB. This patient suffers from X‐linked CGD. The second patient, with autosomal CGD, has a mutation in the donor splice site of intron 1 of CYBA that activates a cryptic donor splice site downstream in intron 1, causing the insertion of intronic sequences in the mRNA. The third patient, recently analyzed, also with autosomal CGD, has a mutation in intron 4 of CYBA, 15 bp from the acceptor splice site. This mutation weakens a branch site and activates a cryptic acceptor splice site, causing the insertion of 14 intronic nucleotides into the mRNA. CONCLUSION: We found three different mutations, one exonic, one in a donor splice site and one intronic, that all caused missplicing of pre‐mRNA. We analyzed these mutations with four different splice prediction programs and found that predictions of splice site strength, splice enhancer and splice silencer protein binding and branch site strength are all essential for correct prediction of pre‐mRNA splicing.
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spelling pubmed-67323212019-09-12 Activation of cryptic splice sites in three patients with chronic granulomatous disease de Boer, Martin van Leeuwen, Karin Hauri‐Hohl, Mathias Roos, Dirk Mol Genet Genomic Med Original Articles BACKGROUND: Chronic granulomatous disease (CGD) is a primary immune deficiency caused by mutations in the genes encoding the structural components of the phagocyte NADPH oxidase. As a result, the patients cannot generate sufficient amounts of reactive oxygen species required for killing pathogenic microorganisms. METHODS: We analyzed NADPH oxidase activity and component expression in neutrophils, performed genomic DNA and cDNA analysis, and used mRNA splicing prediction tools to evaluate the impact of mutations. RESULTS: In two patients with CGD, we had previously found mutations that cause aberrant pre‐mRNA splicing. In one patient an exonic mutation in a cryptic donor splice site caused the deletion of the 3' part of exon 6 from the mRNA of CYBB. This patient suffers from X‐linked CGD. The second patient, with autosomal CGD, has a mutation in the donor splice site of intron 1 of CYBA that activates a cryptic donor splice site downstream in intron 1, causing the insertion of intronic sequences in the mRNA. The third patient, recently analyzed, also with autosomal CGD, has a mutation in intron 4 of CYBA, 15 bp from the acceptor splice site. This mutation weakens a branch site and activates a cryptic acceptor splice site, causing the insertion of 14 intronic nucleotides into the mRNA. CONCLUSION: We found three different mutations, one exonic, one in a donor splice site and one intronic, that all caused missplicing of pre‐mRNA. We analyzed these mutations with four different splice prediction programs and found that predictions of splice site strength, splice enhancer and splice silencer protein binding and branch site strength are all essential for correct prediction of pre‐mRNA splicing. John Wiley and Sons Inc. 2019-07-30 /pmc/articles/PMC6732321/ /pubmed/31364312 http://dx.doi.org/10.1002/mgg3.854 Text en © 2019 Sanquin Blood Supply Organization. Molecular Genetics & Genomic Medicine published by Wiley Periodicals, Inc. This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
spellingShingle Original Articles
de Boer, Martin
van Leeuwen, Karin
Hauri‐Hohl, Mathias
Roos, Dirk
Activation of cryptic splice sites in three patients with chronic granulomatous disease
title Activation of cryptic splice sites in three patients with chronic granulomatous disease
title_full Activation of cryptic splice sites in three patients with chronic granulomatous disease
title_fullStr Activation of cryptic splice sites in three patients with chronic granulomatous disease
title_full_unstemmed Activation of cryptic splice sites in three patients with chronic granulomatous disease
title_short Activation of cryptic splice sites in three patients with chronic granulomatous disease
title_sort activation of cryptic splice sites in three patients with chronic granulomatous disease
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6732321/
https://www.ncbi.nlm.nih.gov/pubmed/31364312
http://dx.doi.org/10.1002/mgg3.854
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