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Nucleic Acids Res. 2015 Mar 31;43(6):3309-17. doi: 10.1093/nar/gkv159. Epub 2015 Mar 3.

Cwc21p promotes the second step conformation of the spliceosome and modulates 3' splice site selection.

Author information

1
Wellcome Trust Centre for Cell Biology, University of Edinburgh, King's Buildings, Mayfield Road, Edinburgh, EH9 3BF, UK.
2
Department of Molecular Genomics, Institute of Molecular Biology of Barcelona (IBMB), 08028 Barcelona, Spain Institució Catalana de Recerca i Estudis Avançats (ICREA), 08010 Barcelona, Spain.
3
Wellcome Trust Centre for Cell Biology, University of Edinburgh, King's Buildings, Mayfield Road, Edinburgh, EH9 3BF, UK jbeggs@ed.ac.uk.

Abstract

Pre-mRNA splicing involves two transesterification steps catalyzed by the spliceosome. How RNA substrates are positioned in each step and the molecular rearrangements involved, remain obscure. Here, we show that mutations in PRP16, PRP8, SNU114 and the U5 snRNA that affect this process interact genetically with CWC21, that encodes the yeast orthologue of the human SR protein, SRm300/SRRM2. Our microarray analysis shows changes in 3' splice site selection at elevated temperature in a subset of introns in cwc21Δ cells. Considering all the available data, we propose a role for Cwc21p positioning the 3' splice site at the transition to the second step conformation of the spliceosome, mediated through its interactions with the U5 snRNP. This suggests a mechanism whereby SRm300/SRRM2, might influence splice site selection in human cells.

PMID:
25740649
PMCID:
PMC4381068
DOI:
10.1093/nar/gkv159
[Indexed for MEDLINE]
Free PMC Article

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