Centre for Plant Molecular Biology (ZMBP), University of Tuebingen, 72076 Tuebingen, Germany.
Chemical Genomics Centre of the Max Planck Society, 44227 Dortmund, Germany.
Plant Cell. 2018 Nov;30(11):2838-2854. doi: 10.1105/tpc.18.00244. Epub 2018 Oct 11.
Introns are removed by the spliceosome, a large macromolecular complex composed of five ribonucleoprotein subcomplexes (U snRNPs). The U1 snRNP, which binds to 5' splice sites, plays an essential role in early steps of the splicing reaction. Here, we show that LETHAL UNLESS CBC7 (LUC7) proteins, which are encoded by a three-member gene family in Arabidopsis, are important for plant development and stress resistance. We show that LUC7 is a U1 snRNP accessory protein by RNA immunoprecipitation experiments and LUC7 protein complex purifications. Transcriptome analyses revealed that LUC7 proteins are not only important for constitutive splicing, but also affect hundreds of alternative splicing events. Interestingly, LUC7 proteins specifically promote splicing of a subset of terminal introns. Splicing of LUC7-dependent introns is a prerequisite for nuclear export, and some splicing events are modulated by stress in a LUC7-dependent manner. Taken together, our results highlight the importance of the U1 snRNP component LUC7 in splicing regulation and suggest a previously unrecognized role of a U1 snRNP accessory factor in terminal intron splicing.
内含子通过剪接体(由五个核糖核蛋白亚基组成的大型大分子复合物)切除。与 5' 剪接位点结合的 U1 snRNP 在剪接反应的早期步骤中发挥着重要作用。在这里,我们发现拟南芥中三个成员基因家族编码的 LETHAL UNLESS CBC7 (LUC7) 蛋白对于植物发育和抗逆性至关重要。我们通过 RNA 免疫沉淀实验和 LUC7 蛋白复合物纯化实验表明,LUC7 是 U1 snRNP 的辅助蛋白。转录组分析表明,LUC7 蛋白不仅对组成性剪接很重要,而且还影响数百个可变剪接事件。有趣的是,LUC7 蛋白特异性促进一组末端内含子的剪接。LUC7 依赖性内含子的剪接是核输出的前提条件,一些剪接事件以 LUC7 依赖的方式受到应激的调节。总之,我们的结果强调了 U1 snRNP 成分 LUC7 在剪接调控中的重要性,并表明 U1 snRNP 辅助因子在末端内含子剪接中具有以前未被认识到的作用。