Cell culture NAIL-MS allows insight into human tRNA and rRNA modification dynamics in vivo

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