Required Elements in tRNA for Methylation by the Eukaryotic tRNA (Guanine-N(2)-) Methyltransferase (Trm11-Trm112 Complex).
Required Elements in tRNA for Methylation by the Eukaryotic tRNA (Guanine-N(2)-) Methyltransferase (Trm11-Trm112 Complex).
复制标题
通过真核tRNA（鸟嘌呤-N（2） - ）甲基转移酶（TRM11-TRM112复合物），tRNA中所需的元素进行甲基化。
DOI：
10.3390/ijms23074046
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发表时间：
2022-04-06
影响因子：
5.6
通讯作者：
Hori H
中科院分区：
文献类型：
--
作者：
Nishida Y;Ohmori S;Kakizono R;Kawai K;Namba M;Okada K;Yamagami R;Hirata A;Hori H
酿酒酵母Trm11和Trm112复合体(Trm11-Trm112)使tRNA中10位鸟苷的2-氨基甲基化，形成N_2-甲基鸟苷。为了确定tRNA中Trm11-Trm112甲基化所需的元件，我们制备了60个tRNA转录变异体，并用Trm11-Trm112检测它们的甲基化。结果表明，前体tRNA不是Trm11-Trm112的底物。此外，CCA末端对于Trm11-Trm112的甲基化是必不可少的，而Trm11-Trm112也只甲基化具有规则大小可变区的tRNA。此外，Trm11-Trm112的甲基化需要G10-C25碱基对。数据还表明，Trm11-Trm112识别反密码子环，而tRNAAla中的U38在甲基化方面起负面作用。同样，tRNACys中的U32-A38碱基对对甲基化产生负面影响。在我们的体外研究中，唯一的例外是tRNAValAAC1。我们的实验表明，tRNAValAAC1转录本被Trm11-Trm112缓慢甲基化。我们从野生型和trm11基因缺失的菌株中纯化了tRNAValAAC1，并证实tRNAValAAC1的一部分在酿酒酵母中被Trm11-Trm112甲基化。因此，我们的研究解释了所有酿酒酵母I类tRNA的m2G10修饰模式，并阐明了Trm11-Trm112结合位点。
The Saccharomyces cerevisiae Trm11 and Trm112 complex (Trm11-Trm112) methylates the 2-amino group of guanosine at position 10 in tRNA and forms N2-methylguanosine. To determine the elements required in tRNA for methylation by Trm11-Trm112, we prepared 60 tRNA transcript variants and tested them for methylation by Trm11-Trm112. The results show that the precursor tRNA is not a substrate for Trm11-Trm112. Furthermore, the CCA terminus is essential for methylation by Trm11-Trm112, and Trm11-Trm112 also only methylates tRNAs with a regular-size variable region. In addition, the G10-C25 base pair is required for methylation by Trm11-Trm112. The data also demonstrated that Trm11-Trm112 recognizes the anticodon-loop and that U38 in tRNAAla acts negatively in terms of methylation. Likewise, the U32-A38 base pair in tRNACys negatively affects methylation. The only exception in our in vitro study was tRNAValAAC1. Our experiments showed that the tRNAValAAC1 transcript was slowly methylated by Trm11-Trm112. However, position 10 in this tRNA was reported to be unmodified G. We purified tRNAValAAC1 from wild-type and trm11 gene deletion strains and confirmed that a portion of tRNAValAAC1 is methylated by Trm11-Trm112 in S. cerevisiae. Thus, our study explains the m2G10 modification pattern of all S. cerevisiae class I tRNAs and elucidates the Trm11-Trm112 binding sites.
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DOI：
10.1261/rna.047639.114
发表时间：
2015-01
期刊：
RNA (New York, N.Y.)
影响因子：
--
作者：
Guy MP;Phizicky EM
通讯作者：
Phizicky EM
DOI：
10.1261/rna.056259.116
发表时间：
2016-09
期刊：
RNA (New York, N.Y.)
影响因子：
--
作者：
Arimbasseri AG;Iben J;Wei FY;Rijal K;Tomizawa K;Hafner M;Maraia RJ
通讯作者：
Maraia RJ
影响因子：
4.5
作者：
Alexandrov, A;Martzen, MR;Phizicky, EM
通讯作者：
Phizicky, EM
影响因子：
4.8
作者：
Awai, Takako;Kimura, Satoshi;Hori, Hiroyuki
通讯作者：
Hori, Hiroyuki
影响因子：
4.5
作者：
Cavaillé, J;Chetouani, F;Bachellerie, JP
通讯作者：
Bachellerie, JP