Issue 5, 2023

Temporal resolution of NAIL-MS of tRNA, rRNA and Poly-A RNA is overcome by actinomycin D

Abstract

RNA is dynamically modified and has the potential to respond to environmental changes and tune translation. The objective of this work is to uncover the temporal limitation of our recently developed cell culture NAIL-MS (nucleic acid isotope labelling coupled mass spectrometry) technology and overcome it. Actinomycin D (AcmD), an inhibitor of transcription, was used in the NAIL-MS context to reveal the origin of hybrid nucleoside signals composed of unlabelled nucleosides and labelled methylation marks. We find that the formation of these hybrid species depends exclusively on transcription for Poly-A RNA and rRNA but is partly transcription-independent for tRNA. This finding suggests that tRNA modifications adapt and are dynamically regulated by cells to overcome e.g. stress. Future studies on the tRNA modification mediated stress response are now accessible and the temporal resolution of NAIL-MS is improved by the use of AcmD.

Graphical abstract: Temporal resolution of NAIL-MS of tRNA, rRNA and Poly-A RNA is overcome by actinomycin D

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Article information

Article type
Paper
Submitted
06 dec 2022
Accepted
15 feb 2023
First published
24 feb 2023
This article is Open Access
Creative Commons BY-NC license

RSC Chem. Biol., 2023,4, 354-362

Temporal resolution of NAIL-MS of tRNA, rRNA and Poly-A RNA is overcome by actinomycin D

F. Hagelskamp, K. Borland, G. Ammann and S. M. Kaiser, RSC Chem. Biol., 2023, 4, 354 DOI: 10.1039/D2CB00243D

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