Yokogawa T, Watanabe Y, Kumazawa Y, Ueda T, Hirao I, Miura K, Watanabe K
Department of Biological Sciences, Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
Nucleic Acids Res. 1991 Nov 25;19(22):6101-5. doi: 10.1093/nar/19.22.6101.
Bovine mitochondrial tRNA(Ser) (UCN) has been thought to have two U-U mismatches at the top of the acceptor stem, as inferred from its gene sequence. However, this unusual structure has not been confirmed at the RNA level. In the course of investigating the structure and function of mitochondrial tRNAs, we have isolated the bovine liver mitochondrial tRNA(Ser) (UCN) and determined its complete sequence including the modified nucleotides. Analysis of the 5'-terminal nucleotide and enzymatic determination of the whole sequence of tRNA(Ser) (UCN) revealed that the tRNA started from the third nucleotide of the putative tRNA(Ser) (UCN) gene, which had formerly been supposed. Enzymatic probing of tRNA(Ser) (UCN) suggests that the tRNA possesses an unusual cloverleaf structure with the following characteristics. (1) There exists only one nucleotide between the acceptor stem with 7 base pairs and the D stem with 4 base pairs. (2) The anticodon stem seems to consist of 6 base pairs. Since the same type of cloverleaf structure as above could be constructed only for mitochondrial tRNA(Ser) (UCN) genes of mammals such as human, rat and mouse, but not for those of non-mammals such as chicken and frog, this unusual secondary structure seems to be conserved only in mammalian mitochondria.
从基因序列推断,牛线粒体tRNA(Ser)(UCN)在受体茎的顶部被认为有两个U-U错配。然而,这种不寻常的结构在RNA水平上尚未得到证实。在研究线粒体tRNA的结构和功能过程中,我们分离出了牛肝线粒体tRNA(Ser)(UCN),并确定了其包括修饰核苷酸在内的完整序列。对tRNA(Ser)(UCN)5'-末端核苷酸的分析以及对其全序列的酶法测定表明,tRNA从先前推测的假定tRNA(Ser)(UCN)基因的第三个核苷酸开始。对tRNA(Ser)(UCN)的酶促探测表明,该tRNA具有一种不寻常的三叶草结构,具有以下特征。(1)在具有7个碱基对的受体茎和具有4个碱基对的D茎之间仅存在一个核苷酸。(2)反密码子茎似乎由6个碱基对组成。由于仅对人类、大鼠和小鼠等哺乳动物的线粒体tRNA(Ser)(UCN)基因可以构建上述相同类型的三叶草结构,而对鸡和青蛙等非哺乳动物的线粒体tRNA(Ser)(UCN)基因则不能构建,这种不寻常的二级结构似乎仅在哺乳动物线粒体中保守。