title: Sequence-specific and Shape-selective RNA Recognition by the Human RNA 5-Methylcytosine Methyltransferase NSun6.
abstract: Human NSun6 is an RNA methyltransferase that catalyzes the transfer of the methyl group from S-adenosyl-l-methionine (SAM) to C72 of tRNAThrand tRNACysIn the current study, we used mass spectrometry to demonstrate that human NSun6 indeed introduces 5-methylcytosine (m5C) into tRNA, as expected. To further reveal the tRNA recognition mechanism of human NSun6, we measured the methylation activity of human NSun6 and its kinetic parameters for different tRNA substrates and their mutants. We showed that human NSun6 requires a well folded, full-length tRNA as its substrate. In the acceptor region, the CCA terminus, the target site C72, the discriminator base U73, and the second and third base pairs (2:71 and 3:70) of the acceptor stem are all important RNA recognition elements for human NSun6. In addition, two specific base pairs (11:24 and 12:23) in the D-stem of the tRNA substrate are involved in interacting with human NSun6. Together, our findings suggest that human NSun6 relies on a delicate network for RNA recognition, which involves both the primary sequence and tertiary structure of tRNA substrates.
authors.affiliation: From the State Key Laboratory of Molecular Biology, CAS Center for Excellence in Molecular Cell Science, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China.|University of the Chinese Academy of Sciences, Beijing 100039, China, and.
authors.forename: Tao
authors.initials: T
authors.lastname: Long
authors.affiliation: From the State Key Laboratory of Molecular Biology, CAS Center for Excellence in Molecular Cell Science, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China.|University of the Chinese Academy of Sciences, Beijing 100039, China, and.
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.affiliation: From the State Key Laboratory of Molecular Biology, CAS Center for Excellence in Molecular Cell Science, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China.|University of the Chinese Academy of Sciences, Beijing 100039, China, and.
authors.forename: Hao
authors.initials: H
authors.lastname: Li
authors.affiliation: From the State Key Laboratory of Molecular Biology, CAS Center for Excellence in Molecular Cell Science, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China.|University of the Chinese Academy of Sciences, Beijing 100039, China, and.
authors.forename: Mi
authors.initials: M
authors.lastname: Zhou
authors.affiliation: From the State Key Laboratory of Molecular Biology, CAS Center for Excellence in Molecular Cell Science, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China.
authors.forename: Xiao-Long
authors.initials: XL
authors.lastname: Zhou
authors.affiliation: From the State Key Laboratory of Molecular Biology, CAS Center for Excellence in Molecular Cell Science, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China, liurj@sibcb.ac.cn.
authors.forename: Ru-Juan
authors.initials: RJ
authors.lastname: Liu
authors.affiliation: From the State Key Laboratory of Molecular Biology, CAS Center for Excellence in Molecular Cell Science, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China, edwang@sibcb.ac.cn.|University of the Chinese Academy of Sciences, Beijing 100039, China, and.|the School of Life Science and Technology, ShanghaiTech University, 319 Yue Yang Road, Shanghai 200031, China.
authors.forename: En-Duo
authors.initials: ED
authors.lastname: Wang
mesh_terms: D006801:Humans; D008745:Methylation; D000072417:Protein Domains; D017510:Protein Folding; D012363:RNA, Transfer, Thr / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012359:tRNA Methyltransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D012363:RNA, Transfer, Thr; C000602638:NSUN6 protein, human; D012359:tRNA Methyltransferases
keywords: 5-methylcytosine; NSun family; RNA methylation; RNA methyltransferase; S-adenosylmethionine (SAM); methyltransferase; substrate recognition; tRNA modification; transfer RNA (tRNA)