{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T07:17:04Z","timestamp":1791962224020,"version":"4.3.4"},"reference-count":39,"publisher":"Oxford University Press (OUP)","issue":"14","license":[{"start":{"date-parts":[[2021,7,16]],"date-time":"2021-07-16T00:00:00Z","timestamp":1626393600000},"content-version":"vor","delay-in-days":1,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31670801"],"award-info":[{"award-number":["31670801"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31822015"],"award-info":[{"award-number":["31822015"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81870896"],"award-info":[{"award-number":["81870896"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Shanghai Key Laboratory of Embryo Original Diseases","award":["Shelab201904"],"award-info":[{"award-number":["Shelab201904"]}]},{"name":"Key Laboratory of Reproductive Genetics, Ministry of Education, Zhejiang University","award":["ZDFY2020-RG-0003"],"award-info":[{"award-number":["ZDFY2020-RG-0003"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2021,8,20]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>tRNAs harbor the most diverse posttranscriptional modifications. The 3-methylcytidine (m3C) is widely distributed at position C32 (m3C32) of eukaryotic tRNAThr and tRNASer species. m3C32 is decorated by the single methyltransferase Trm140 in budding yeasts; however, two (Trm140 and Trm141 in fission yeasts) or three enzymes (METTL2A, METTL2B and METTL6 in mammals) are involved in its biogenesis. The rationale for the existence of multiple m3C32 methyltransferases and their substrate discrimination mechanism is hitherto unknown. Here, we revealed that both METTL2A and METTL2B are expressed in vivo. We purified human METTL2A, METTL2B, and METTL6 to high homogeneity. We successfully reconstituted m3C32 modification activity for tRNAThr by METT2A and for tRNASer(GCU) by METTL6, assisted by seryl-tRNA synthetase (SerRS) in vitro. Compared with METTL2A, METTL2B exhibited dramatically lower activity in vitro. Both G35 and t6A at position 37 (t6A37) are necessary but insufficient prerequisites for tRNAThr m3C32 formation, while the anticodon loop and the long variable arm, but not t6A37, are key determinants for tRNASer(GCU) m3C32 biogenesis, likely being recognized synergistically by METTL6 and SerRS, respectively. Finally, we proposed a mutually exclusive substrate selection model to ensure correct discrimination among multiple tRNAs by multiple m3C32 methyltransferases.<\/jats:p>","DOI":"10.1093\/nar\/gkab603","type":"journal-article","created":{"date-parts":[[2021,7,1]],"date-time":"2021-07-01T19:50:13Z","timestamp":1625169013000},"page":"8309-8323","source":"Crossref","is-referenced-by-count":62,"title":["Mutually exclusive substrate selection strategy by human m3C RNA transferases METTL2A and METTL6"],"prefix":"10.1093","volume":"49","author":[{"given":"Xue-Ling","family":"Mao","sequence":"first","affiliation":[{"name":"State Key Laboratory of Molecular Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zi-Han","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Molecular Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meng-Han","family":"Huang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Molecular Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jin-Tao","family":"Wang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Molecular Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing-Bo","family":"Zhou","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Molecular Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qing-Run","family":"Li","sequence":"additional","affiliation":[{"name":"CAS Key Laboratory of Systems Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hong","family":"Xu","sequence":"additional","affiliation":[{"name":"Shanghai Key Laboratory of Embryo Original Diseases, the International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, 910 Heng Shan Road, Shanghai 200030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xi-Jin","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Neurology, Xinhua Hospital, School of Medicine, Shanghai Jiao Tong University, 1665 Kong Jiang Road, Shanghai 200092, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3701-5965","authenticated-orcid":false,"given":"Xiao-Long","family":"Zhou","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Molecular Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2021,7,15]]},"reference":[{"key":"2021081818125413300_B1","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1146\/annurev-genet-110711-155641","article-title":"Biosynthesis and function of posttranscriptional modifications of transfer RNAs","volume":"46","author":"El\u00a0Yacoubi","year":"2012","journal-title":"Annu. Rev. Genet."},{"key":"2021081818125413300_B2","doi-asserted-by":"crossref","first-page":"e1595","DOI":"10.1002\/wrna.1595","article-title":"Naturally occurring modified ribonucleosides","volume":"11","author":"McCown","year":"2020","journal-title":"Wiley Interdiscip Rev RNA"},{"key":"2021081818125413300_B3","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1038\/s41580-021-00342-0","article-title":"The expanding world of tRNA modifications and their disease relevance","volume":"22","author":"Suzuki","year":"2021","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"2021081818125413300_B4","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/bs.enz.2017.03.005","article-title":"The importance of being modified: The role of RNA modifications in translational fidelity","volume":"41","author":"Agris","year":"2017","journal-title":"Enzymes"},{"key":"2021081818125413300_B5","doi-asserted-by":"crossref","first-page":"1771","DOI":"10.1261\/rna.056531.116","article-title":"tRNA base methylation identification and quantification via high-throughput sequencing","volume":"22","author":"Clark","year":"2016","journal-title":"RNA"},{"key":"2021081818125413300_B6","doi-asserted-by":"crossref","first-page":"1100","DOI":"10.1261\/rna.2652611","article-title":"A domain of the actin binding protein Abp140 is the yeast methyltransferase responsible for 3-methylcytidine modification in the tRNA anti-codon loop","volume":"17","author":"D'Silva","year":"2011","journal-title":"RNA"},{"key":"2021081818125413300_B7","doi-asserted-by":"crossref","first-page":"1111","DOI":"10.1261\/rna.2653411","article-title":"Actin-binding protein ABP140 is a methyltransferase for 3-methylcytidine at position 32 of tRNAs in Saccharomyces cerevisiae","volume":"17","author":"Noma","year":"2011","journal-title":"RNA"},{"key":"2021081818125413300_B8","doi-asserted-by":"crossref","first-page":"14695","DOI":"10.1074\/jbc.M117.798298","article-title":"Three distinct 3-methylcytidine (m3C) methyltransferases modify tRNA and mRNA in mice and humans","volume":"292","author":"Xu","year":"2017","journal-title":"J. Biol. Chem."},{"key":"2021081818125413300_B9","doi-asserted-by":"crossref","first-page":"406","DOI":"10.1261\/rna.059667.116","article-title":"S. cerevisiae Trm140 has two recognition modes for 3-methylcytidine modification of the anticodon loop of tRNA substrates","volume":"23","author":"Han","year":"2017","journal-title":"RNA"},{"key":"2021081818125413300_B10","doi-asserted-by":"crossref","first-page":"1400","DOI":"10.1261\/rna.056259.116","article-title":"Evolving specificity of tRNA 3-methyl-cytidine-32 (m3C32) modification: a subset of tRNAsSer requires N6-isopentenylation of A37","volume":"22","author":"Arimbasseri","year":"2016","journal-title":"RNA"},{"key":"2021081818125413300_B11","doi-asserted-by":"crossref","first-page":"2510","DOI":"10.1038\/s41467-020-16321-6","article-title":"DALRD3 encodes a protein mutated in epileptic encephalopathy that targets arginine tRNAs for 3-methylcytosine modification","volume":"11","author":"Lentini","year":"2020","journal-title":"Nat. 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China Life Sci."},{"key":"2021081818125413300_B30","doi-asserted-by":"crossref","first-page":"1529","DOI":"10.4161\/15476286.2014.992277","article-title":"Diversity of the biosynthesis pathway for threonylcarbamoyladenosine (t6A), a universal modification of tRNA","volume":"11","author":"Thiaville","year":"2014","journal-title":"RNA Biol."},{"key":"2021081818125413300_B31","first-page":"10534","article-title":"SerRS-tRNASec complex structures reveal mechanism of the first step in selenocysteine biosynthesis","volume":"43","author":"Wang","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"2021081818125413300_B32","doi-asserted-by":"crossref","first-page":"1404","DOI":"10.1126\/science.8128220","article-title":"The 2.9 A crystal structure of T. thermophilus seryl-tRNA synthetase complexed with tRNASer","volume":"263","author":"Biou","year":"1994","journal-title":"Science"},{"key":"2021081818125413300_B33","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1261\/rna.035808.112","article-title":"tRNAHis 5-methylcytidine levels increase in response to several growth arrest conditions in Saccharomyces cerevisiae","volume":"19","author":"Preston","year":"2013","journal-title":"RNA"},{"key":"2021081818125413300_B34","doi-asserted-by":"crossref","first-page":"12833","DOI":"10.1093\/nar\/gkaa1123","article-title":"Bacterial tRNA 2\u2032-O-methylation is dynamically regulated under stress conditions and modulates innate immune response","volume":"48","author":"Galvanin","year":"2020","journal-title":"Nucleic Acids Res."},{"key":"2021081818125413300_B35","doi-asserted-by":"crossref","first-page":"eaaz4551","DOI":"10.1126\/sciadv.aaz4551","article-title":"METTL6 is a tRNA m3C methyltransferase that regulates pluripotency and tumor cell growth","volume":"6","author":"Ignatova","year":"2020","journal-title":"Sci. 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