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

SUBMITTER: Reichle VF 

PROVIDER: S-EPMC6582434 | biostudies-literature | 2018 Dec

REPOSITORIES: biostudies-literature