Laubinger Sascha, Sachsenberg Timo, Zeller Georg, Busch Wolfgang, Lohmann Jan U, Rätsch Gunnar, Weigel Detlef
Department of Molecular Biology, Max Planck Institute for Developmental Biology, 72076 Tübingen, Germany.
Proc Natl Acad Sci U S A. 2008 Jun 24;105(25):8795-800. doi: 10.1073/pnas.0802493105. Epub 2008 Jun 12.
The processing of Arabidopsis thaliana microRNAs (miRNAs) from longer primary transcripts (pri-miRNAs) requires the activity of several proteins, including DICER-LIKE1 (DCL1), the double-stranded RNA-binding protein HYPONASTIC LEAVES1 (HYL1), and the zinc finger protein SERRATE (SE). It has been noted before that the morphological appearance of weak se mutants is reminiscent of plants with mutations in ABH1/CBP80 and CBP20, which encode the two subunits of the nuclear cap-binding complex. We report that, like SE, the cap-binding complex is necessary for proper processing of pri-miRNAs. Inactivation of either ABH1/CBP80 or CBP20 results in decreased levels of mature miRNAs accompanied by apparent stabilization of pri-miRNAs. Whole-genome tiling array analyses reveal that se, abh1/cbp80, and cbp20 mutants also share similar splicing defects, leading to the accumulation of many partially spliced transcripts. This is unlikely to be an indirect consequence of improper miRNA processing or other mRNA turnover pathways, because introns retained in se, abh1/cbp80, and cbp20 mutants are not affected by mutations in other genes required for miRNA processing or for nonsense-mediated mRNA decay. Taken together, our results uncover dual roles in splicing and miRNA processing that distinguish SE and the cap-binding complex from specialized miRNA processing factors such as DCL1 and HYL1.
拟南芥微小RNA(miRNA)从较长的初级转录本(pri-miRNA)加工而来需要几种蛋白质的参与,包括Dicer样蛋白1(DCL1)、双链RNA结合蛋白下弯叶片1(HYL1)和锌指蛋白锯齿状(SE)。之前已经有人指出,弱se突变体的形态外观让人联想到在ABH1/CBP80和CBP20中发生突变的植物,这两个基因编码核帽结合复合体的两个亚基。我们报道,与SE一样,帽结合复合体对于pri-miRNA的正确加工也是必需的。ABH1/CBP80或CBP20的失活会导致成熟miRNA水平降低,同时pri-miRNA明显稳定。全基因组平铺阵列分析表明,se、abh1/cbp80和cbp20突变体也存在相似的剪接缺陷,导致许多部分剪接的转录本积累。这不太可能是miRNA加工不当或其他mRNA周转途径的间接后果,因为se、abh1/cbp80和cbp20突变体中保留的内含子不受miRNA加工或无义介导的mRNA降解所需的其他基因中的突变影响。综上所述,我们的结果揭示了SE和帽结合复合体在剪接和miRNA加工中的双重作用,这将它们与DCL1和HYL这样的专门miRNA加工因子区分开来。