THUMPD3-TRMT112 is a m2G methyltransferase working on a broad range of tRNA substrates.
THUMPD3-TRMT112 is a m2G methyltransferase working on a broad range of tRNA substrates.
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THUMPD3-TRMT112 是一种 m2G 甲基转移酶，作用于多种 tRNA 底物。
DOI：
10.1093/nar/gkab927
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发表时间：
2021-11-18
影响因子：
14.9
通讯作者：
Liu RJ
中科院分区：
文献类型：
--
作者：
Yang WQ;Xiong QP;Ge JY;Li H;Zhu WY;Nie Y;Lin X;Lv D;Li J;Lin H;Liu RJ
转录后修饰影响tRNA生物学，并与人类疾病密切相关。然而，由于识别修饰酶的困难，后生动物中tRNA修饰的功能分析进展缓慢。例如，真核生物中普遍存在的位于tRNAs第六位的N_2-甲基鸟苷(M2G)的生物发生和功能长期以来一直是个谜。在这里，我们使用反向遗传学和RNA-MS相结合的方法，确定了THUMPD3(THUMPD3)是人类细胞中tRNA：m2G6形成的原因。然而，THUMPD3单独不能修饰tRNA。相反，多功能甲基转移酶亚单位TRM112样蛋白(TRMT112)与THUMPD3相互作用，激活其甲基转移酶活性。在体外酶分析系统中，THUMPD3-TRMT112通过识别成熟tRNA的特征3‘-CCA，能够甲基化所有26个含有G6的人胞质tRNA。我们还证明了tRNATrp的m2G7是由THUMPD3-TRMT112引入的。此外，THUMPD3在小鼠组织中广泛表达，在睾丸中的表达水平极高。THUMPD3基因敲除的细胞表现出全球蛋白质合成受损和生长减慢。我们的数据突出了tRNA：m2G6/7修饰的重要性，并为进一步研究m2G在精子tRNA衍生片段中的作用铺平了道路。
Post-transcriptional modifications affect tRNA biology and are closely associated with human diseases. However, progress on the functional analysis of tRNA modifications in metazoans has been slow because of the difficulty in identifying modifying enzymes. For example, the biogenesis and function of the prevalent N2-methylguanosine (m2G) at the sixth position of tRNAs in eukaryotes has long remained enigmatic. Herein, using a reverse genetics approach coupled with RNA-mass spectrometry, we identified that THUMP domain-containing protein 3 (THUMPD3) is responsible for tRNA: m2G6 formation in human cells. However, THUMPD3 alone could not modify tRNAs. Instead, multifunctional methyltransferase subunit TRM112-like protein (TRMT112) interacts with THUMPD3 to activate its methyltransferase activity. In the in vitro enzymatic assay system, THUMPD3–TRMT112 could methylate all the 26 tested G6-containing human cytoplasmic tRNAs by recognizing the characteristic 3′-CCA of mature tRNAs. We also showed that m2G7 of tRNATrp was introduced by THUMPD3–TRMT112. Furthermore, THUMPD3 is widely expressed in mouse tissues, with an extremely high level in the testis. THUMPD3-knockout cells exhibited impaired global protein synthesis and reduced growth. Our data highlight the significance of the tRNA: m2G6/7 modification and pave a way for further studies of the role of m2G in sperm tRNA derived fragments.
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影响因子：
14.9
作者：
Chan PP;Lowe TM
通讯作者：
Lowe TM
影响因子：
4.8
作者：
Awai, Takako;Kimura, Satoshi;Hori, Hiroyuki
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Hori, Hiroyuki
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作者：
EDQVIST, J;BLOMQVIST, K;STRABY, KB
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STRABY, KB
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4.5
作者：
Freund, Isabel;Buhl, Daniel K.;Eigenbrod, Tatjana
通讯作者：
Eigenbrod, Tatjana
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4.8
作者：
Armengaud, J;Urbonavicius, J;Grosjean, H
通讯作者：
Grosjean, H