Handa H, Kubo N, Kadowaki K
Laboratory of Plant Genecology, Hokkaido National Agricultural Experiment Station, Sapporo, Japan.
Mol Gen Genet. 1998 May;258(3):199-207. doi: 10.1007/s004380050723.
Here we describe the structure and expression of the mitochondrial gene that encodes the S4 subunit of the mitochondrial ribosome (rps4) in two higher plants, rapeseed and rice. The ORFs of rapeseed and rice rps4 genes are 362 and 352 codons long, respectively, and their corresponding transcripts are edited (C-to-U conversions) in 19 and 15 positions, respectively. The amino acid sequences deduced from the sequences of the two rps4 genes are 78.1% identical to each other and show about 50% identity to the liverwort (Marchantia polymorpha) mitochondrial RPS4 polypeptide. Both higher plant mitochondrial genes have an inserted sequence (of 137 and 141 codons for rapeseed and rice, respectively) relative to the liverwort rps4 gene. In rice rps4, the 3' portion of the inserted sequence can potentially form two stem-loop structures that resemble two domains characteristic of group II introns, i.e., domains V and VI. However, RT-PCR analysis showed that these sequences were not spliced out during maturation of the rps4 transcripts. Antibodies specific for the plant RPS4 proteins were prepared. Protein blot analysis revealed that RPS4 polypeptides are present in both rapeseed and rice mitochondria, whose sizes are in agreement with the molecular weights calculated from the non-spliced sequences. The rps4 genes and RPS4 polypeptides have been also identified in the mitochondria of wheat, maize, soybean, and turnip by Southern and Western analyses, respectively.
在此,我们描述了编码线粒体核糖体S4亚基(rps4)的线粒体基因在两种高等植物油菜和水稻中的结构与表达。油菜和水稻rps4基因的开放阅读框分别长362和352个密码子,其相应的转录本分别在19个和15个位置发生编辑(C到U的转换)。从这两个rps4基因序列推导的氨基酸序列彼此间有78.1%的同一性,并且与地钱(多歧紫菜)线粒体RPS4多肽有约50%的同一性。相对于地钱rps4基因,这两个高等植物线粒体基因都有一个插入序列(油菜和水稻分别为137和141个密码子)。在水稻rps4中,插入序列的3'部分可能形成两个茎环结构,类似于II类内含子的两个特征结构域,即结构域V和结构域VI。然而,逆转录-聚合酶链反应(RT-PCR)分析表明,这些序列在rps4转录本成熟过程中并未被剪接出去。制备了针对植物RPS4蛋白的特异性抗体。蛋白质印迹分析显示,RPS4多肽存在于油菜和水稻的线粒体中,其大小与根据未剪接序列计算的分子量一致。通过Southern分析和Western分析,分别在小麦、玉米、大豆和芜菁的线粒体中也鉴定出了rps4基因和RPS4多肽。