Control of Pre-mRNA Splicing by the General Splicing Factors PUF60 and U2AF65

Hastings, M. L., Allemand, E., Duelli, D. M., Myers, M. P., Krainer, A. R. (June 2007) Control of Pre-mRNA Splicing by the General Splicing Factors PUF60 and U2AF65. PLoS ONE, 2 (6). e538. ISSN 1932-6203 (Electronic)

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URL: http://www.ncbi.nlm.nih.gov/pubmed/17579712
DOI: 10.1371/journal.pone.0000538

Abstract

Pre-mRNA splicing is a crucial step in gene expression, and accurate recognition of splice sites is an essential part of this process. Splice sites with weak matches to the consensus sequences are common, though it is not clear how such sites are efficiently utilized. Using an in vitro splicing-complementation approach, we identified PUF60 as a factor that promotes splicing of an intron with a weak 3' splice-site. PUF60 has homology to U2AF(65), a general splicing factor that facilitates 3' splice-site recognition at the early stages of spliceosome assembly. We demonstrate that PUF60 can functionally substitute for U2AF(65)in vitro, but splicing is strongly stimulated by the presence of both proteins. Reduction of either PUF60 or U2AF(65) in cells alters the splicing pattern of endogenous transcripts, consistent with the idea that regulation of PUF60 and U2AF(65) levels can dictate alternative splicing patterns. Our results indicate that recognition of 3' splice sites involves different U2AF-like molecules, and that modulation of these general splicing factors can have profound effects on splicing.

Item Type: Paper
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > introns > intron splicing
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > pre-mRNA
CSHL Authors:
Communities: CSHL labs > Krainer lab
Depositing User: CSHL Librarian
Date: 20 June 2007
Date Deposited: 15 Nov 2011 15:09
Last Modified: 09 Apr 2014 14:17
PMCID: PMC1888729
Related URLs:
URI: https://repository.cshl.edu/id/eprint/23028

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