Identification of the yeast gene encoding the tRNA m1G methyltransferase responsible for modification at position 9
Identification of the yeast gene encoding the tRNA m1G methyltransferase responsible for modification at position 9
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DOI：
10.1261/rna.5070303
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发表时间：
2003-05-01
期刊：
影响因子：
4.5
通讯作者：
Phizicky, EM
中科院分区：
文献类型：
--
作者：
Jackman, JE;Montange, RK;Phizicky, EM
tRNA在鸟苷N-1位甲基化形成m（1）G在自然界中广泛存在。它发生在来自所有三个王国的tRNA的37位，并且催化该反应的甲基转移酶从其他人的先前工作中已知对于大肠杆菌和酵母酿酒酵母中的细胞生长至关重要。m（1）G也广泛存在于真核tRNA的9位，但相应的甲基转移酶未知。我们已经使用了一个生物化学基因组学的方法与纯化的酵母菌的集合。GST-ORF融合蛋白显示m（1）G（9）形成的酵母tRNA（Gly）与ORF YOL 093 w相关，命名为TRM 10。缺乏Trm 10 p的提取物在体内对大多数（如果不是所有的话）m（1）G（9）修饰的tRNA负责，基于两个结果：从trm 10-delta/trm 10-delta染色纯化的tRNA（Gly）缺乏可检测的m（1）G;并且对于所有9个m（1）G（9），在trm 10-δ/trm 10-δ衍生的tRNA中去除了在m（1）G（9）处发生的引物延伸阻断。包含可通过这种方法测试的物种。trm 10-delta/trm 10-delta菌株无明显生长检测。Trm 10 p与酵母m（1）G（37）甲基转移酶Trm 5 p或其直系同源物没有可检测的相似性。Trm 10 p同源物广泛存在于真核生物和许多古细菌中，在几种后生动物中有多个同源物，包括人类中的至少三个。
Methylation of tRNA at the N-1 position of guanosine to form m(1)G occurs widely in nature. It occurs at position 37 in tRNAs from all three kingdoms, and the methyltransferase that catalyzes this reaction is known from previous work of others to be critically important for cell growth in Escherichia coli and the yeast Saccharomyces cerevisiae. m(1)G is also widely found at position 9 in eukaryotic tRNAs, but the corresponding methyltransferase was unknown. We have used a biochemical genomics approach with a collection of purified yeast. GST-ORF fusion proteins to show that m(1)G(9) formation of yeast tRNA(Gly) is associated with ORF YOL093w, named TRM10. Extracts lacking Trm 10p is responsible in vivo for most if not all m(1)G(9) modification tRNAs, based on two results: tRNA(Gly) purified from a trm 10-delta/trm 10-delta stain is lacking detectable m(1)G; and a primer extension block occurring at m(1)G(9) is removed in trm 10-delta/trm 10-delta-derived tRNAs for all 9 m(1)G(9)-containing species that were testable by this method. There is no obvious growth detect of trm 10-delta/trm 10-delta stains. Trm 10p bears no detectable resemblance to the yeast m(1)G(37) methyltransferase, Trm5p, or its orthologs. Trm 10p homologs are found widely in eukaryotes and many archaea, with multiple homologs in several metazoans, including at least three in humans.