Functional splicing assay shows a pathogenic intronic mutation in neurofibromatosis type 1 (NF1) due to intronic sequence exonization
Functional splicing assay shows a pathogenic intronic mutation in neurofibromatosis type 1 (NF1) due to intronic sequence exonization
Genomic variations with no apparent effect ("neutral polymorphisms") may have a significant effect on splicing. The effect of this type of mutation is difficult to spot, unless a functional assay is undertaken. In our study, DNA sequencing of a patient with clinically defined neurofibromatosis type 1 (NF1) showed only a single polymorphism in intron 30 due to an A>G transition 279 nucleotides from the 3' splice site. Using a minigene splicing assay we conclusively show that this change produces a cryptic exon with a 3' SS defined by the nucleotide change and the unexpected activation of a very weak 5'SS. Further site directed mutagenesis studies aimed at identifying the signals involved in the cryptic exon inclusion were carried out. Interestingly we find that particular characteristics of the cryptic 5' SS are essential for its inclusion. Significantly an additional single nucleotide change disrupting the cryptic 5'ss consensus sequence rescues the effect of the pathogenetic mutation resulting in normal splicing.
294-295
Raponi, M.
80317d2e-fc3d-4350-bebd-fd978837f7b1
Upadhyaya, M.
2e8fdcf8-f5d1-491b-b315-55db22432e1b
Baralle, D.
faac16e5-7928-4801-9811-8b3a9ea4bb91
March 2006
Raponi, M.
80317d2e-fc3d-4350-bebd-fd978837f7b1
Upadhyaya, M.
2e8fdcf8-f5d1-491b-b315-55db22432e1b
Baralle, D.
faac16e5-7928-4801-9811-8b3a9ea4bb91
Raponi, M., Upadhyaya, M. and Baralle, D.
(2006)
Functional splicing assay shows a pathogenic intronic mutation in neurofibromatosis type 1 (NF1) due to intronic sequence exonization.
Human Mutation, 27 (3), .
(doi:10.1002/humu.9412).
Abstract
Genomic variations with no apparent effect ("neutral polymorphisms") may have a significant effect on splicing. The effect of this type of mutation is difficult to spot, unless a functional assay is undertaken. In our study, DNA sequencing of a patient with clinically defined neurofibromatosis type 1 (NF1) showed only a single polymorphism in intron 30 due to an A>G transition 279 nucleotides from the 3' splice site. Using a minigene splicing assay we conclusively show that this change produces a cryptic exon with a 3' SS defined by the nucleotide change and the unexpected activation of a very weak 5'SS. Further site directed mutagenesis studies aimed at identifying the signals involved in the cryptic exon inclusion were carried out. Interestingly we find that particular characteristics of the cryptic 5' SS are essential for its inclusion. Significantly an additional single nucleotide change disrupting the cryptic 5'ss consensus sequence rescues the effect of the pathogenetic mutation resulting in normal splicing.
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Published date: March 2006
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Local EPrints ID: 48496
URI: http://eprints.soton.ac.uk/id/eprint/48496
ISSN: 1059-7794
PURE UUID: 730dae89-29a7-4864-b351-0a46e310920a
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Date deposited: 26 Sep 2007
Last modified: 16 Mar 2024 03:57
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Author:
M. Raponi
Author:
M. Upadhyaya
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