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| Subject Categories: RNA |
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| The EMBO Journal
(2006) 25, 3813–3822, doi:10.1038/sj.emboj.7601258 Published online 3 August 2006 |
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| Mutation in the U2 snRNA influences exon interactions of U5 snRNA loop 1 during pre-mRNA splicing |
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| Joanne C McGrail, Elaine M Tatum1 and Raymond T O'Keefe |
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Faculty of Life Sciences, The University of Manchester, Manchester, UK To whom correspondence should be addressed Raymond T O'Keefe, Faculty of Life Sciences, The University of Manchester, Michael Smith Building, Oxford Road, Manchester M13 9PT, UK. Tel.: +44 161 275 7393; Fax: +44 161 275 5082; E-mail: rokeefe@manchester.ac.uk 1 Present address: Chiron Corporation, Cowley, Uxbridge UB8 2AD, UK Received 14 February 2006; Accepted 4 July 2006; Published online 3 August 2006. |
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| Abstract |
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| The U2 and U6 snRNAs contribute to the catalysis of intron removal while U5 snRNA loop 1 holds the exons for ligation during pre-mRNA splicing. It is unclear how different exons are positioned precisely with U5 loop 1. Here, we investigate the role of U2 and U6 in positioning the exons with U5 loop 1. Reconstitution in vitro of spliceosomes with mutations in U2 allows U5–pre-mRNA interactions before the first step of splicing. However, insertion in U2 helix Ia disrupts U5–exon interactions with the intron lariat-3' exon splicing intermediate. Conversely, U6 helix Ia insertions prevent U5–pre-mRNA interactions before the first step of splicing. In vivo, synthetic lethal interactions have been identified between U2 insertion and U5 loop 1 insertion mutants. Additionally, analysis of U2 insertion mutants in vivo reveals that they influence the efficiency, but not the accuracy of splicing. Our data suggest that U2 aligns the exons with U5 loop 1 for ligation during the second step of pre-mRNA splicing. |
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| Keywords: pre-mRNA splicing, spliceosome, U2 snRNA, U5 snRNA, yeast |
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Top of page MORE ARTICLES LIKE THISThese links to content published by NPG are automatically generated NEWS AND VIEWSThe spliceosome shows its metal Nature Structural Biology News and Views (01 Jan 2001) Nature News and Views (29 Dec 1988) RESEARCHThe EMBO Journal Article (15 Feb 1997) Functional analysis of the U5 snRNA loop 1 in the second catalytic step of yeast pre-mRNA splicing The EMBO Journal Article (15 Jan 1998) |
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