Ohta M, Sugita M, Sugiura M
Center for Gene Research, Nagoya University, Japan.
Plant Mol Biol. 1995 Feb;27(3):529-39. doi: 10.1007/BF00019319.
Five chloroplast RNA-binding proteins with consensus sequence-type RNA-binding domains have been isolated from tobacco chloroplasts. Here we report three nuclear genes for similar chloroplast RNA-binding proteins (cp29, cp31 and cp33) from Arabidopsis thaliana. Each of the three genes consists of four exons and three introns and their exon/intron junctions were determined by sequencing respective cDNAs. In vitro import assays showed that all three proteins are located in chloroplasts. The three genes are singly-copy each and the transcription start sites were determined to be 80/82 bp (cp29) and 76/88 bp (cp31) upstream from the translational initiation codons. Northern blot analysis revealed that the three genes are transcribed both in leaves and roots, but the transcript level in leaves is higher than in roots. Phylogenetic analysis of chloroplast RNA-binding proteins so far identified shows that these proteins can be classified into three groups. Tobacco and Arabidopsis have these three types of proteins and structural features of each group are conserved between the two plants, suggesting that they are important for chloroplast functions. Interestingly, cp31 (238 amino acids) shares the identical amino acid sequence from the 30th to the last (238th) residues (including two RNA-binding domains) with the Arabidopsis nucleolin-like ribonucleoprotein, FMV3bp [11]. FMV3bp lacks a transit-peptide and must be located in the nucleus or the cytoplasm.
已从烟草叶绿体中分离出五种具有共有序列型RNA结合结构域的叶绿体RNA结合蛋白。本文我们报道了来自拟南芥的三个与叶绿体RNA结合蛋白(cp29、cp31和cp33)相似的核基因。这三个基因均由四个外显子和三个内含子组成,通过对各自cDNA进行测序确定了它们的外显子/内含子连接。体外导入实验表明这三种蛋白均定位于叶绿体中。这三个基因均为单拷贝,转录起始位点确定为位于翻译起始密码子上游80/82 bp(cp29)和76/88 bp(cp31)处。Northern印迹分析表明这三个基因在叶和根中均有转录,但叶中的转录水平高于根。对目前已鉴定的叶绿体RNA结合蛋白的系统发育分析表明,这些蛋白可分为三组。烟草和拟南芥都有这三种类型的蛋白,且两组植物中每组的结构特征均保守,这表明它们对叶绿体功能很重要。有趣的是,cp31(238个氨基酸)与拟南芥核仁样核糖核蛋白FMV3bp从第30个残基到最后一个(第238个)残基(包括两个RNA结合结构域)具有相同的氨基酸序列[11]。FMV3bp缺乏转运肽,必定定位于细胞核或细胞质中。