Journal
RNA
Volume 7, Issue 6, Pages 785-792Publisher
COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT
DOI: 10.1017/S1355838201010263
Keywords
exonic splicing enhancer; nematodes; SR proteins; trans-splicing; U2AF
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Funding
- NIAID NIH HHS [AI-28799] Funding Source: Medline
- NIA NIH HHS [AG0001015-15] Funding Source: Medline
- NIGMS NIH HHS [GM 31528] Funding Source: Medline
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Trans-splicing requires that 5' and 3' splice sites be independently recognized. Here, we have used mutational analyses and a sensitive nuclease protection assay to determine the mechanism of trans-3' splice site recognition in vitro. Efficient recognition of the 3' splice site is dependent upon both the sequence of the 3' splice site itself and enhancer elements located in the 3' exon. We show that the presence of three distinct classes of enhancers results in increased binding of U2 snRNP to the branchpoint region. Several lines of evidence strongly suggest that the increased binding of U2 snRNP is mediated by USAF. These results expand the roles of enhancers in constitutive splicing and provide direct support for the recruitment model of enhancer function.
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