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NAIL-MS reveals the repair of 2-methylthiocytidine by AlkB in E. coli.

Abstract：

:RNAs contain post-transcriptional modifications, which fulfill a variety of functions in translation, secondary structure stabilization and cellular stress survival. Here, 2-methylthiocytidine (ms2C) is identified in tRNA of E. coli and P. aeruginosa using NAIL-MS (nucleic acid isotope labeling coupled mass spectrometry) in combination with genetic screening experiments. ms2C is only found in 2-thiocytidine (s2C) containing tRNAs, namely tRNAArgCCG, tRNAArgICG, tRNAArgUCU and tRNASerGCU at low abundances. ms2C is not formed by commonly known tRNA methyltransferases. Instead, we observe its formation in vitro and in vivo during exposure to methylating agents. More than half of the s2C containing tRNA can be methylated to carry ms2C. With a pulse-chase NAIL-MS experiment, the repair mechanism by AlkB dependent sulfur demethylation is demonstrated in vivo. Overall, we describe ms2C as a bacterial tRNA modification and damage product. Its repair by AlkB and other pathways is demonstrated in vivo by our powerful NAIL-MS approach.

journal_name

Nat Commun

journal_title

Nature communications

authors

Reichle VF,Petrov DP,Weber V,Jung K,Kellner S

doi

10.1038/s41467-019-13565-9

subject

Has Abstract

pub_date

2019-12-06 00:00:00

pages

5600

issue

1

issn

2041-1723

pii

10.1038/s41467-019-13565-9

journal_volume

10

pub_type

杂志文章

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