Kwak Kyung Jin, Kim Bo Mi, Lee Kwanuk, Kang Hunseung
Department of Plant Biotechnology, College of Agriculture and Life Sciences, Chonnam National University, Gwangju 500-757, Korea.
J Exp Bot. 2017 May 17;68(11):2731-2739. doi: 10.1093/jxb/erx138.
Despite increasing understanding of the importance of the splicing of U12-type introns in plant development, the key question of which U12 intron-containing genes are essential for plant development has not yet been explored. Here, we assessed the functional role of the quatre-quart1 (QQT1) gene, one of the ~230 U12 intron-containing genes in Arabidopsis thaliana. Expression of QQT1 in the U11/U12-31K small nuclear ribonucleoprotein mutant (31k) rescued the developmental-defect phenotypes of the 31k mutant, whereas the miRNA-mediated qqt1 knockdown mutants displayed severe defects in growth and development, including severely arrested stem growth, small size, and the formation of serrated leaves. The structures of the shoot apical meristems in the qqt1 mutants were abnormal and disordered. Identification of QQT1-interacting proteins via a yeast two-hybrid screening and a firefly luciferase complementation-imaging assay revealed that a variety of proteins, including many chloroplast-targeted proteins, interacted with QQT1. Importantly, the levels of chloroplast-targeted proteins in the chloroplast were reduced, and the chloroplast structure was abnormal in the qqt1 mutant. Collectively, these results provide clear evidence that QQT1 is an indispensable U12 intron-containing gene whose correct splicing is crucial for the normal development of Arabidopsis.
尽管人们对U12型内含子剪接在植物发育中的重要性的认识不断加深,但哪些含U12内含子的基因对植物发育至关重要这一关键问题尚未得到探索。在这里,我们评估了拟南芥中约230个含U12内含子的基因之一——四分之一四分体1(QQT1)基因的功能作用。QQT1在U11/U12 - 31K小核核糖核蛋白突变体(31k)中的表达挽救了31k突变体的发育缺陷表型,而miRNA介导的qqt1基因敲低突变体在生长和发育方面表现出严重缺陷,包括茎生长严重停滞、植株矮小以及锯齿状叶片的形成。qqt1突变体的茎尖分生组织结构异常且紊乱。通过酵母双杂交筛选和萤火虫荧光素酶互补成像分析鉴定与QQT1相互作用的蛋白质,结果显示多种蛋白质,包括许多靶向叶绿体的蛋白质,与QQT1相互作用。重要的是,qqt1突变体中靶向叶绿体的蛋白质在叶绿体中的水平降低,并且叶绿体结构异常。总体而言,这些结果提供了明确的证据,表明QQT1是一个不可或缺的含U12内含子的基因,其正确剪接对于拟南芥的正常发育至关重要。