Cell Culture NAIL-MS Allows Insight into Human TRNA and RRNA Modification Dynamics in Vivo

Overview

Journal Nat Commun

Specialty Biology

Date 2021 Jan 16

PMID 33452242

Citations 14

Authors

Matthias Heiss

Felix Hagelskamp

Virginie Marchand

Yuri Motorin

Stefanie Kellner

Affiliations

Soon will be listed here.

Abstract

Recently, studies about RNA modification dynamics in human RNAs are among the most controversially discussed. As a main reason, we identified the unavailability of a technique which allows the investigation of the temporal processing of RNA transcripts. Here, we present nucleic acid isotope labeling coupled mass spectrometry (NAIL-MS) for efficient, monoisotopic stable isotope labeling in both RNA and DNA in standard cell culture. We design pulse chase experiments and study the temporal placement of modified nucleosides in tRNA and 18S rRNA. In existing RNAs, we observe a time-dependent constant loss of modified nucleosides which is masked by post-transcriptional methylation mechanisms and thus undetectable without NAIL-MS. During alkylation stress, NAIL-MS reveals an adaptation of tRNA modifications in new transcripts but not existing ones. Overall, we present a fast and reliable stable isotope labeling strategy which allows in-depth study of RNA modification dynamics in human cell culture.

Citing Articles

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