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Nucleic Acids Res. 2011 Mar;39(5):1868-79. doi: 10.1093/nar/gkq1042. Epub 2010 Nov 9.

Analysis of in situ pre-mRNA targets of human splicing factor SF1 reveals a function in alternative splicing.

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  • 1Department of Cell Biology, Faculty of Sciences, University of Geneva, 30 quai Ernest-Ansermet, CH-1211 Geneva.

Abstract

The conserved pre-mRNA splicing factor SF1 is implicated in 3' splice site recognition by binding directly to the intron branch site. However, because SF1 is not essential for constitutive splicing, its role in pre-mRNA processing has remained mysterious. Here, we used crosslinking and immunoprecipitation (CLIP) to analyze short RNAs directly bound by human SF1 in vivo. SF1 bound mainly pre-mRNAs, with 77% of target sites in introns. Binding to target RNAs in vitro was dependent on the newly defined SF1 binding motif ACUNAC, strongly resembling human branch sites. Surprisingly, the majority of SF1 binding sites did not map to the expected position near 3' splice sites. Instead, target sites were distributed throughout introns, and a smaller but significant fraction occurred in exons within coding and untranslated regions. These data suggest a more complex role for SF1 in splicing regulation. Indeed, SF1 silencing affected alternative splicing of endogenous transcripts, establishing a previously unexpected role for SF1 and branch site-like sequences in splice site selection.

PMID:
21062807
[PubMed - indexed for MEDLINE]
PMCID:
PMC3061054
Free PMC Article

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