Menand B, Maréchal-Drouard L, Sakamoto W, Dietrich A, Wintz H
Institut de Biologie Moléculaire des Plantes/Centre National de la Recherche Scientifique, Université Louis Pasteur, 12 rue du Général Zimmer, F-67084 Strasbourg Cedex, France.
Proc Natl Acad Sci U S A. 1998 Sep 1;95(18):11014-9. doi: 10.1073/pnas.95.18.11014.
One-fifth of the tRNAs used in plant mitochondrial translation is coded for by chloroplast-derived tRNA genes. To understand how aminoacyl-tRNA synthetases have adapted to the presence of these tRNAs in mitochondria, we have cloned an Arabidopsis thaliana cDNA coding for a methionyl-tRNA synthetase. This enzyme was chosen because chloroplast-like elongator tRNAMet genes have been described in several plant species, including A. thaliana. We demonstrate here that the isolated cDNA codes for both the chloroplastic and the mitochondrial methionyl-tRNA synthetase (MetRS). The protein is transported into isolated chloroplasts and mitochondria and is processed to its mature form in both organelles. Transient expression assays using the green fluorescent protein demonstrated that the N-terminal region of the MetRS is sufficient to address the protein to both chloroplasts and mitochondria. Moreover, characterization of MetRS activities from mitochondria and chloroplasts of pea showed that only one MetRS activity exists in each organelle and that both are indistinguishable by their behavior on ion exchange and hydrophobic chromatographies. The high degree of sequence similarity between A. thaliana and Synechocystis MetRS strongly suggests that the A. thaliana MetRS gene described here is of chloroplast origin.
植物线粒体翻译中使用的五分之一的tRNA由叶绿体来源的tRNA基因编码。为了了解氨酰-tRNA合成酶如何适应线粒体中这些tRNA的存在,我们克隆了拟南芥中一个编码甲硫氨酰-tRNA合成酶的cDNA。选择这种酶是因为在包括拟南芥在内的几种植物中已经描述了叶绿体样延伸因子tRNAMet基因。我们在此证明,分离的cDNA编码叶绿体和线粒体甲硫氨酰-tRNA合成酶(MetRS)。该蛋白质被转运到分离的叶绿体和线粒体中,并在这两种细胞器中加工成成熟形式。使用绿色荧光蛋白的瞬时表达分析表明,MetRS的N端区域足以将该蛋白质靶向叶绿体和线粒体。此外,对豌豆线粒体和叶绿体中MetRS活性的表征表明,每个细胞器中仅存在一种MetRS活性,并且它们在离子交换和疏水色谱上的行为无法区分。拟南芥和集胞藻MetRS之间高度的序列相似性强烈表明,这里描述的拟南芥MetRS基因起源于叶绿体。