Low U1 snRNP dependence at the NF1 exon 29 donor site
Low U1 snRNP dependence at the NF1 exon 29 donor site
Many disease-causing splicing mutations described in the literature produce changes in splice sites (SS) or in exon-regulatory sequences. The delineation of these splice aberrations can provide important insights into novel regulation mechanisms. In this study, we evaluated the effect of patient variations in neurofibromatosis type 1 (NF1) exon 29 and its 5'SS surrounding area on its splicing process. Only two of all nonsense, missense, synonymous and intronic variations analyzed in this study clearly altered exon 29 inclusion/exclusion levels. In particular, the intronic mutation +5g>a had the strongest effect, resulting in total exon exclusion. This finding prompted us to evaluate the exon 29 5'SS in relation to its ability to bind U1 snRNP. This was performed by direct analysis of the ability of U1 to bind to wild-type and mutant donor sites, by engineering an in vitro splicing system to directly evaluate the functional importance of U1 snRNA base pairing with the exon 29 donor site, and by coexpression of mutant U1 snRNP molecules to try to rescue exon 29 inclusion in vivo. The results revealed a low dependency on the presence of U1 snRNP, and suggest that exon 29 donor site definition may depend on alternative mechanisms of 5'SS recognition
2060-2073
Raponi, Michael
f465e77f-b9bf-4c32-80d6-43c0787542b9
Buratti, Emanuele
57e8e002-a8c2-409a-ac29-2fc4a1d1c8b9
Dassie, Elisa
18566ce2-048b-47b8-928b-1c2baccf149a
Upadhyaya, Meena
63533b81-2307-4ea0-8980-6b3abd5b055b
Baralle, Diane
faac16e5-7928-4801-9811-8b3a9ea4bb91
April 2009
Raponi, Michael
f465e77f-b9bf-4c32-80d6-43c0787542b9
Buratti, Emanuele
57e8e002-a8c2-409a-ac29-2fc4a1d1c8b9
Dassie, Elisa
18566ce2-048b-47b8-928b-1c2baccf149a
Upadhyaya, Meena
63533b81-2307-4ea0-8980-6b3abd5b055b
Baralle, Diane
faac16e5-7928-4801-9811-8b3a9ea4bb91
Raponi, Michael, Buratti, Emanuele, Dassie, Elisa, Upadhyaya, Meena and Baralle, Diane
(2009)
Low U1 snRNP dependence at the NF1 exon 29 donor site.
Febs Journal, 276 (7), .
(doi:10.1111/j.1742-4658.2009.06941.x).
Abstract
Many disease-causing splicing mutations described in the literature produce changes in splice sites (SS) or in exon-regulatory sequences. The delineation of these splice aberrations can provide important insights into novel regulation mechanisms. In this study, we evaluated the effect of patient variations in neurofibromatosis type 1 (NF1) exon 29 and its 5'SS surrounding area on its splicing process. Only two of all nonsense, missense, synonymous and intronic variations analyzed in this study clearly altered exon 29 inclusion/exclusion levels. In particular, the intronic mutation +5g>a had the strongest effect, resulting in total exon exclusion. This finding prompted us to evaluate the exon 29 5'SS in relation to its ability to bind U1 snRNP. This was performed by direct analysis of the ability of U1 to bind to wild-type and mutant donor sites, by engineering an in vitro splicing system to directly evaluate the functional importance of U1 snRNA base pairing with the exon 29 donor site, and by coexpression of mutant U1 snRNP molecules to try to rescue exon 29 inclusion in vivo. The results revealed a low dependency on the presence of U1 snRNP, and suggest that exon 29 donor site definition may depend on alternative mechanisms of 5'SS recognition
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Published date: April 2009
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Local EPrints ID: 69634
URI: http://eprints.soton.ac.uk/id/eprint/69634
ISSN: 1742-464X
PURE UUID: 9021ef1e-74d1-4cdc-a073-c6e11d5ceb8f
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Date deposited: 25 Nov 2009
Last modified: 14 Mar 2024 02:53
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Author:
Michael Raponi
Author:
Emanuele Buratti
Author:
Elisa Dassie
Author:
Meena Upadhyaya
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