NAIL‐MS in E. coli Determines the Source and Fate of Methylation in tRNA

Abstract: In all domains of life, the nucleobases of tRNA can be methylated. These methylations are introduced either by enzymes or by the reaction of methylating agents with the nucleophilic centers of the nucleobases. Herein, we present a systematic approach to identify the methylation sites within RNA in vitro and in vivo. For discrimination between enzymatic tRNA methylation and tRNA methylation damage in bacteria, we used nucleic acid isotope labeling coupled mass spectrometry (NAIL‐MS). With NAIL‐MS, we clearly observed the formation of 7‐methylguanosine, 3‐methyluridine, and 6‐methyladenosine during exposure of bacteria to the alkylating agent methyl methanesulfonate (MMS) in vivo. These damage products were not reported to form in tRNA in vivo, as they were masked by the enzymatically formed modified nucleosides in previous studies. In addition, we found formation of the known damage products 1‐methyladenosine and 3‐methylcytidine in vivo. With a dynamic NAIL‐MS setup, we observed tRNA repair by demethylation of these two RNA modifications in vivo. Furthermore, we saw the potential repair of 6‐methyladenosine but not 7‐methylguanosine in bacterial tRNA.

Object Details

• Location: Deutsche Nationalbibliothek Frankfurt am Main
• Extent: Online-Ressource
• Language: Englisch

References and Relationships

• Bibliographic citation: NAIL‐MS in E. coli Determines the Source and Fate of Methylation in tRNA ; volume:19 ; number:24 ; year:2018 ; pages:2575-2583 ; extent:9 ; ChemBioChem ; 19, Heft 24 (2018), 2575-2583 (gesamt 9)

Contributors, Places and Time

• Creator: Reichle, Valentin F. ; Weber, Verena ; Kaiser, Stefanie

Further information

• DOI: 10.1002/cbic.201800525
• URN: urn:nbn:de:101:1-2022082305540825916757
• Rights: Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
• Last update: 15.08.2025, 7:26 AM CEST