Rokov-Plavec Jasmina, Dulic Morana, Duchêne Anne-Marie, Weygand-Durasevic Ivana
Department of Chemistry, Faculty of Natural Sciences, University of Zagreb, Horvatovac 102a, 10000 Zagreb, Croatia.
Plant Cell Rep. 2008 Jul;27(7):1157-68. doi: 10.1007/s00299-008-0542-9. Epub 2008 Apr 5.
Aminoacyl-tRNA synthetases (AARSs) play a critical role in translation and are thus required in three plant protein-synthesizing compartments: cytosol, mitochondria and plastids. A systematic study had previously shown extensive sharing of organellar AARSs from Arabidopsis thaliana, mostly between mitochondria and chloroplasts. However, distribution of AARSs from monocot species, such as maize, has never been experimentally investigated. Here we demonstrate dual targeting of maize seryl-tRNA synthetase, SerZMo, into both mitochondria and chloroplasts using combination of complementary methods, including in vitro import assay, transient expression analysis of green fluorescent protein (GFP) fusions and immunodetection. We also show that SerZMo dual localization is established by the virtue of an ambiguous targeting peptide. Full-length SerZMo protein fused to GFP is targeted to chloroplast stromules, indicating that SerZMo protein performs its function in plastid stroma. The deletion mutant lacking N-terminal region of the ambiguous SerZMo targeting peptide was neither targeted into mitochondria nor chloroplasts, indicating the importance of this region in both mitochondrial and chloroplastic import.
氨酰-tRNA合成酶(AARSs)在翻译过程中发挥着关键作用,因此在植物的三个蛋白质合成区室中都有需求:细胞质、线粒体和质体。此前一项系统性研究表明,拟南芥的细胞器AARSs存在广泛共享,主要是在线粒体和叶绿体之间。然而,从未对单子叶植物物种(如玉米)的AARSs分布进行过实验研究。在这里,我们使用包括体外导入分析、绿色荧光蛋白(GFP)融合蛋白的瞬时表达分析和免疫检测等互补方法相结合,证明了玉米丝氨酰-tRNA合成酶SerZMo可双重靶向线粒体和叶绿体。我们还表明,SerZMo的双重定位是由一个模糊的靶向肽实现的。与GFP融合的全长SerZMo蛋白靶向叶绿体的基质小管,表明SerZMo蛋白在质体基质中发挥其功能。缺失模糊的SerZMo靶向肽N端区域的缺失突变体既不靶向线粒体也不靶向叶绿体,表明该区域在向线粒体和叶绿体的导入过程中都很重要。