biostudies-literatureSun YDeutsche ForschungsgemeinschaftChinese Scholarship CouncilKlaus Tschira Foundation11197-11212https://www.ebi.ac.uk/biostudies/studies/S-EPMC10639084biostudies-literatureUnknown51(20)Queuosine (Q) is a complex tRNA modification found in bacteria and eukaryotes at position 34 of four tRNAs with a GUN anticodon, and it regulates the translational efficiency and fidelity of the respective codons that differ at the Wobble position. In bacteria, the biosynthesis of Q involves two precursors, preQ0 and preQ1, whereas eukaryotes directly obtain Q from bacterial sources. The study of queuosine has been challenging due to the limited availability of high-throughput methods for its detection and analysis. Here, we have employed direct RNA sequencing using nanopore technology to detect the modification of tRNAs with Q and Q precursors. These modifications were detected with high accuracy on synthetic tRNAs as well as on tRNAs extracted from Schizosaccharomyces pombe and EscherichNucleic acids researchDetection of queuosine and queuosine precursors in tRNAs by direct RNA sequencing.PMC10639084EH237/ 13-2SPP178400.013.2021EH237/ 19-1Naarmann-de Vries IPiechotta MDieterich CSun YEhrenhofer-Murray AEfalseDetection of queuosine and queuosine precursors in tRNAs by direct RNA sequencing.Queuosine (Q) is a complex tRNA modification found in bacteria and eukaryotes at position 34 of four tRNAs with a GUN anticodon, and it regulates the translational efficiency and fidelity of the respective codons that differ at the Wobble position. In bacteria, the biosynthesis of Q involves two precursors, preQ0 and preQ1, whereas eukaryotes directly obtain Q from bacterial sources. The study of queuosine has been challenging due to the limited availability of high-throughput methods for its detection and analysis. Here, we have employed direct RNA sequencing using nanopore technology to detect the modification of tRNAs with Q and Q precursors. These modifications were detected with high accuracy on synthetic tRNAs as well as on tRNAs extracted from Schizosaccharomyces pombe and Escherich2023-01-01T00:00:00Z2023 Nov2026-06-23T03:20:39.082Z2025-04-04T13:15:41.742ZS-EPMC106390843781187210.1093/nar/gkad826