control of pre-mrna splicing by the general splicing factors puf60 and u2af65控制pre-mrna通用剪接因子puf60和u2af65拼接.pdfVIP

Control of Pre-mRNA Splicing by the General Splicing Factors PUF60 and U2AF65 ¤ Michelle L. Hastings, Eric Allemand , Dominik M. Duelli, Michael P. Myers, Adrian R. Krainer* Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, United States of America 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 39 splice-site. PUF60 has homology to U2AF65, a general splicing factor that facilitates 3 9 splice-site recognition at the early stages of spliceosome assembly. We demonstrate that PUF60 can functionally substitute for U2AF65 in vitro, but splicing is strongly stimulated by the presence of both proteins. Reduction of either PUF60 or U2AF65 in cells alters the splicing pattern of endogenous transcripts, consistent with the idea that regulation of PUF60 and U2AF65 levels can dictate alternative splicing patterns. Our results indicate that recognition of 39 splice sites involves different U2AF-like molecules, and that modulation of these general splicing factors can have profound effects on splicing. Citation: Hastings ML, Allemand E, Duelli DM, Myers MP, Krainer AR (2007) Control of Pre-mRNA Splicing by the General Splicing Factors PUF60 and U2AF65. PLoS ONE 2(6): e538. doi:10.1371/journal.pone.0000538 INTRODUCTION are many cases of introns with weak splice sites that are Accurate pre-mRNA splicing is essential for proper gene constitutively spliced, and examples