State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
Plant Physiol. 2013 Jan;161(1):165-78. doi: 10.1104/pp.112.206706. Epub 2012 Nov 6.
In prokaryotic and eukaryotic cells, the 3'-5'-exonucleolytic decay and processing of RNAs are essential for RNA metabolism. However, the understanding of the mechanism of 3'-5'-exonucleolytic decay in plants is very limited. Here, we report the characterization of an Arabidopsis (Arabidopsis thaliana) transfer DNA insertional mutant that shows severe growth defects in early seedling growth, including delayed germination and cotyledon expansion, thinner yellow/pale-green leaves, and a slower growth rate. High-efficiency thermal asymmetric interlaced polymerase chain reaction analysis showed that the insertional locus was in the sixth exon of AT4G27490, encoding a predicted 3'-5'-exonuclease, that contained a conserved RNase phosphorolytic domain with high similarity to RRP41, designated RRP41L. Interestingly, we detected highly accumulated messenger RNAs (mRNAs) that encode seed storage protein and abscisic acid (ABA) biosynthesis and signaling pathway-related protein during the early growth stage in rrp41l mutants. The mRNA decay kinetics analysis for seed storage proteins, 9-cis-epoxycarotenoid dioxygenases, and ABA INSENSITIVEs revealed that RRP41L catalyzed the decay of these mRNAs in the cytoplasm. Consistent with these results, the rrp41l mutant was more sensitive to ABA in germination and root growth than wild-type plants, whereas overexpression lines of RRP41L were more resistant to ABA in germination and root growth than wild-type plants. RRP41L was localized to both the cytoplasm and nucleus, and RRP41L was preferentially expressed in seedlings. Altogether, our results showed that RRP41L plays an important role in seed germination and early seedling growth by mediating specific cytoplasmic mRNA decay in Arabidopsis.
在原核和真核细胞中,RNA 的 3'-5'-外切核酸酶降解和加工对于 RNA 代谢至关重要。然而,植物中 3'-5'-外切核酸酶降解的机制理解非常有限。在这里,我们报道了一个拟南芥(Arabidopsis thaliana)转座子插入突变体的特征,该突变体在早期幼苗生长中表现出严重的生长缺陷,包括发芽延迟和子叶扩张、叶片更薄、更黄/淡绿色以及生长速度较慢。高效热不对称交错聚合酶链反应分析表明,插入位点位于 AT4G27490 的第六外显子中,该基因编码一个预测的 3'-5'-外切核酸酶,该酶含有一个与 RRP41 高度相似的保守核糖核酸酶磷酸化结构域,命名为 RRP41L。有趣的是,我们在 rrp41l 突变体中检测到在早期生长阶段高度积累的信使 RNA(mRNA),这些 mRNA 编码种子贮藏蛋白和脱落酸(ABA)生物合成和信号通路相关蛋白。种子贮藏蛋白、9-顺式-环氧类胡萝卜素双加氧酶和 ABA 不敏感蛋白的 mRNA 衰减动力学分析表明,RRP41L 催化这些 mRNA 在细胞质中的衰减。与这些结果一致,rrp41l 突变体在发芽和根生长中对 ABA 比野生型植物更敏感,而 RRP41L 的过表达系在发芽和根生长中对 ABA 比野生型植物更具抗性。RRP41L 定位于细胞质和细胞核,RRP41L 优先在幼苗中表达。总之,我们的结果表明,RRP41L 通过介导特定的细胞质 mRNA 衰减在拟南芥中在种子发芽和早期幼苗生长中发挥重要作用。