Department of Gene Expression, Adam Mickiewicz University, Poznan, Poland.
Nucleic Acids Res. 2010 Jan;38(1):265-78. doi: 10.1093/nar/gkp869. Epub 2009 Oct 28.
The nuclear cap-binding protein complex (CBC) participates in 5' splice site selection of introns that are proximal to the mRNA cap. However, it is not known whether CBC has a role in alternative splicing. Using an RT-PCR alternative splicing panel, we analysed 435 alternative splicing events in Arabidopsis thaliana genes, encoding mainly transcription factors, splicing factors and stress-related proteins. Splicing profiles were determined in wild type plants, the cbp20 and cbp80(abh1) single mutants and the cbp20/80 double mutant. The alternative splicing events included alternative 5' and 3' splice site selection, exon skipping and intron retention. Significant changes in the ratios of alternative splicing isoforms were found in 101 genes. Of these, 41% were common to all three CBC mutants and 15% were observed only in the double mutant. The cbp80(abh1) and cbp20/80 mutants had many more changes in alternative splicing in common than did cbp20 and cbp20/80 suggesting that CBP80 plays a more significant role in alternative splicing than CBP20, probably being a platform for interactions with other splicing factors. Cap-binding proteins and the CBC are therefore directly involved in alternative splicing of some Arabidopsis genes and in most cases influenced alternative splicing of the first intron, particularly at the 5' splice site.
核帽结合蛋白复合物 (CBC) 参与靠近 mRNA 帽的内含子 5' 剪接位点的选择。然而,CBC 是否在可变剪接中起作用尚不清楚。使用 RT-PCR 可变剪接面板,我们分析了拟南芥基因中的 435 个可变剪接事件,这些基因主要编码转录因子、剪接因子和应激相关蛋白。在野生型植物、cbp20 和 cbp80(abh1) 单突变体和 cbp20/80 双突变体中确定了剪接谱。可变剪接事件包括 5' 和 3' 剪接位点选择、外显子跳跃和内含子保留的可变剪接。在 101 个基因中发现了可变剪接同工型比率的显著变化。其中,41%在所有三个 CBC 突变体中都相同,15%仅在双突变体中观察到。Cbp80(abh1) 和 cbp20/80 突变体之间的可变剪接变化比 cbp20 和 cbp20/80 之间的变化更为常见,这表明 CBP80 在可变剪接中比 CBP20 发挥更重要的作用,可能是与其他剪接因子相互作用的平台。因此,帽结合蛋白和 CBC 直接参与了一些拟南芥基因的可变剪接,并且在大多数情况下影响了第一个内含子的可变剪接,特别是在 5' 剪接位点。