Biology Division, National Cancer Center Research Institute, Tsukiji 5-1, Chuo-ku, Tokyo 104, Japan.
EMBO J. 1986 Jun;5(6):1307-11. doi: 10.1002/j.1460-2075.1986.tb04360.x.
The three major glutamine tRNAs of Tetrahymena thermophila were isolated and their nucleotide sequences determined by post-labeling techniques. Two of these tRNAs show unusual codon recognition: a previously isolated tRNA(UmUA) and a second species with CUA in the anticodon (tRNA(CUA)). These two tRNAs recognize two of the three termination codons on natural mRNAs in a reticulocyte system. tRNA(UmUA) reads the UAA codon of alpha-globin mRNA and the UAG codon of tobacco mosaic virus (TMV) RNA, whereas tRNA(CUA) recognizes only UAG. This indicates that Tetrahymena uses UAA and UAG as glutamine codons and that UGA may be the only functional termination codon. A notable feature of these two tRNAs is their unusually strong readthrough efficiency, e.g. purified tRNA(CUA) achieves complete readthrough over the UAG stop codon of TMV RNA. The third major tRNA of Tetrahymena has a UmUG anticodon and presumably reads the two normal glutamine codons CAA and CAG. The sequence homology between tRNA(UmUG) and tRNA(UmUA) is 81%, whereas that between tRNA(CUA) and tRNA(UmUA) is 95%, indicating that the two unusual tRNAs evolved from the normal tRNA early in ciliate evolution. Possible events leading to an altered genetic code in ciliates are discussed.
嗜热四膜虫的三种主要谷氨酰胺 tRNA 已被分离出来,并通过标记后技术确定了它们的核苷酸序列。其中两种 tRNA 显示出不寻常的密码子识别:一种是先前分离的 tRNA(UmUA),另一种是反密码子中含有 CUA 的第二种物种(tRNA(CUA))。这两种 tRNA 在网织红细胞系统中识别天然 mRNA 上的三个终止密码子中的两个。tRNA(UmUA)读取 alpha-球蛋白 mRNA 的 UAA 密码子和烟草花叶病毒 (TMV) RNA 的 UAG 密码子,而 tRNA(CUA)仅识别 UAG。这表明嗜热四膜虫将 UAA 和 UAG 用作谷氨酰胺密码子,而 UGA 可能是唯一的功能终止密码子。这两种 tRNA 的一个显著特点是它们异常强的通读效率,例如,纯化的 tRNA(CUA)在 TMV RNA 的 UAG 终止密码子上实现完全通读。嗜热四膜虫的第三种主要 tRNA 具有 UmUG 反密码子,推测可以读取两个正常的谷氨酰胺密码子 CAA 和 CAG。tRNA(UmUG)与 tRNA(UmUA)之间的序列同源性为 81%,而 tRNA(CUA)与 tRNA(UmUA)之间的序列同源性为 95%,表明这两种不寻常的 tRNA 是在纤毛动物进化的早期从正常 tRNA 进化而来的。还讨论了纤毛动物中遗传密码发生改变的可能事件。