TY - JOUR AU - Guo, Wei AU - Kaczmarczyk, Igor AU - Kopietz, Kevin AU - Flegler, Florian AU - Russo, Stefano AU - Cigirgan, Ege AU - Chramiec-Głąbik, Andrzej AU - Koziej, Łukasz AU - Cirzi, Cansu AU - Peschek, Jirka AU - Reuter, Klaus AU - Helm, Mark AU - Glatt, Sebastian AU - Tuorto, Francesca TI - Queuosine is incorporated into precursor tRNA before splicing. JO - Nature Communications VL - 16 IS - 1 SN - 2041-1723 CY - [London] PB - Springer Nature M1 - DKFZ-2025-01594 SP - 7044 PY - 2025 N1 - DKFZ-ZMBH Alliance AB - Each newly transcribed tRNA molecule must undergo processing and receive modifications to become functional. Queuosine (Q) is a tRNA modification present at position 34 of four tRNAs with 'GUN' anticodons. Among these, the precursor of tRNATyr carries an intronic sequence within the anticodon loop that is removed by an essential non-canonical splicing event. The functional and temporal coupling between tRNA-splicing and Q-incorporation remains elusive. Here, we demonstrate in vitro and in vivo that intron-containing precursors of tRNATyr are modified with Q or with the Q-derivative galactosyl-queuosine (galQ) before being spliced. We show that this order of events is conserved in mouse, human, flies and worms. Using single particle cryo-EM, we confirm that pre-tRNATyr is a bona fide substrate of the QTRT1/2 complex, which catalyzes the incorporation of Q into the tRNA. Our results elucidate the hierarchical interplay that coordinates Q-incorporation and splicing in eukaryotic tRNAs, providing a relevant but unappreciated aspect of the cellular tRNA maturation process. KW - Animals KW - RNA Splicing KW - Nucleoside Q: metabolism KW - Humans KW - Mice KW - RNA Precursors: metabolism KW - RNA Precursors: genetics KW - RNA, Transfer: metabolism KW - RNA, Transfer: genetics KW - Cryoelectron Microscopy KW - Introns: genetics KW - Anticodon: genetics KW - Caenorhabditis elegans: genetics KW - Pentosyltransferases: metabolism KW - Nucleoside Q (NLM Chemicals) KW - RNA Precursors (NLM Chemicals) KW - RNA, Transfer (NLM Chemicals) KW - Anticodon (NLM Chemicals) KW - Pentosyltransferases (NLM Chemicals) LB - PUB:(DE-HGF)16 C6 - pmid:40745156 DO - DOI:10.1038/s41467-025-62220-z UR - https://inrepo02.dkfz.de/record/303249 ER -