a combinatorial code for splicing silencing uagg and gggg motifs拼接的组合代码沉默uagg段勇图案.pdfVIP

Open access, freely available online PLoS BIOLOGY A Combinatorial Code for Splicing Silencing: UAGG and GGGG Motifs 1¤a[ 2¤b[ 1 3 1* Kyoungha Han , Gene Yeo , Ping An , Christopher B. Burge , Paula J. Grabowski 1 Department of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America, 2 Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Boston, Massachusetts, United States of America, 3 Department of Biology, Massachusetts Institute of Technology, Boston, Massachusetts, United States of America Alternative pre-mRNA splicing is widely used to regulate gene expression by tuning the levels of tissue-specific mRNA isoforms. Few regulatory mechanisms are understood at the level of combinatorial control despite numerous sequences, distinct from splice sites, that have been shown to play roles in splicing enhancement or silencing. Here we use molecular approaches to identify a ternary combination of exonic UAGG and 59-splice-site-proximal GGGG motifs that functions cooperatively to silence the brain-region-specific CI cassette exon (exon 19) of the glutamate NMDA R1 receptor (GRIN1) transcript. Disruption of three components of the motif pattern converted the CI cassette into a constitutive exon, while predominant skipping was conferred when the same components were introduced, de novo, into a heterologous constitutive exon. Predominant exon silencing was directed by the motif pattern in the presence of six competing exonic splicing enhancers, and this effect was retained after systematically repositioning the two exonic UAGGs within the CI cassette. In this system, hnRNP A1 was shown to mediate silencing whil