Department of Plant Biotechnology and Agricultural Plant Stress Research Center, College of Agriculture and Life Sciences, Chonam National University, Gwangju 500-757, Korea.
Plant Cell. 2010 Dec;22(12):3951-62. doi: 10.1105/tpc.110.079103. Epub 2010 Dec 10.
U12 introns are removed from precursor-mRNA by a U12 intron-specific spliceosome that contains U11 and U12 small nuclear ribonucleoproteins. Although several proteins unique to the U12-type spliceosome have been identified, the manner by which they affect U12-dependent intron splicing as well as plant growth and development remain largely unknown. Here, we assessed the role of U11/U12-31K, a U12-type spliceosomal protein in Arabidopsis thaliana. T-DNA-tagged homozygote lines for U11/U12-31K could not be obtained, and heterozygote mutants were defective for seed maturation, indicating that U11/U12-31K is essential for the normal development of Arabidopsis. Knockdown of U11/U12-31K by artificial microRNA caused a defect in proper U12 intron splicing, resulting in abnormal stem growth and development of Arabidopsis. This defect in proper splicing was not restricted to specific U12-type introns, but most U12 intron splicing was influenced by U11/U12-31K. The stunted inflorescence stem growth was recovered by exogenously applied gibberellic acid (GA), but not by cytokinin, auxin, or brassinosteroid. GA metabolism-related genes were highly downregulated in U11/U12-31K knockdown plants. Importantly, U11/U12-31K was determined to harbor RNA chaperone activity. We propose that U11/U12-31K is an RNA chaperone that is indispensible for proper U12 intron splicing and for normal growth and development of plants.
U12 内含子通过含有 U11 和 U12 小核核糖核蛋白的 U12 内含子特异性剪接体从前体-mRNA 中被切除。尽管已经鉴定出几种特有的 U12 剪接体的蛋白质,但它们影响 U12 依赖性内含子剪接以及植物生长和发育的方式在很大程度上仍然未知。在这里,我们评估了拟南芥中 U11/U12-31K(一种 U12 剪接体蛋白)的作用。不能获得 T-DNA 标记的 U11/U12-31K 纯合子系,杂合突变体在种子成熟方面有缺陷,表明 U11/U12-31K 对于拟南芥的正常发育是必不可少的。人工 microRNA 敲低 U11/U12-31K 导致 U12 内含子剪接异常,导致拟南芥茎生长和发育异常。这种适当剪接的缺陷不仅限于特定的 U12 型内含子,而是大多数 U12 内含子剪接受到 U11/U12-31K 的影响。外源性赤霉素(GA)可以恢复短缩的花序茎生长,但细胞分裂素、生长素或油菜素内酯不能恢复。U11/U12-31K 敲低植物中 GA 代谢相关基因高度下调。重要的是,U11/U12-31K 被确定具有 RNA 伴侣活性。我们提出,U11/U12-31K 是一种 RNA 伴侣,对于 U12 内含子剪接的适当进行以及植物的正常生长和发育是必不可少的。