Department of Materials Science and Biotechnology, Graduate School of Science and Engineering, Ehime University, Bunkyo 3, Matsuyama, Ehime 790-8577, Japan.
FEBS Lett. 2013 Nov 1;587(21):3575-80. doi: 10.1016/j.febslet.2013.09.021. Epub 2013 Sep 25.
Thermoplasma acidophilum is a thermo-acidophilic archaeon. We purified tRNA(Leu) (UAG) from T. acidophilum using a solid-phase DNA probe method and determined the RNA sequence after determining via nucleoside analysis and m(7)G-specific aniline cleavage because it has been reported that T. acidophilum tRNA contains m(7)G, which is generally not found in archaeal tRNAs. RNA sequencing and liquid chromatography-mass spectrometry revealed that the m(7)G modification exists at a novel position 49. Furthermore, we found several distinct modifications, which have not previously been found in archaeal tRNA, such as 4-thiouridine9, archaeosine13 and 5-carbamoylmethyuridine34. The related tRNA modification enzymes and their genes are discussed.
嗜酸热原体是一种嗜热嗜酸古菌。我们使用固相 DNA 探针法从嗜酸热原体中纯化 tRNA(Leu)(UAG)，并通过核苷分析和 m(7)G 特异性苯胺裂解来确定 RNA 序列，因为据报道，嗜酸热原体 tRNA 含有 m(7)G，通常在古菌 tRNA 中不存在。RNA 测序和液相色谱-质谱分析表明，m(7)G 修饰存在于一个新的位置 49。此外，我们还发现了一些以前在古菌 tRNA 中未发现的独特修饰，如 4-硫尿嘧啶 9、假尿嘧啶 13 和 5-碳氨甲基尿嘧啶 34。讨论了相关的 tRNA 修饰酶及其基因。
