Queuosine is incorporated into precursor tRNA before splicing

Wei Guo, Igor Kaczmarczyk, Kevin Kopietz, Florian Flegler, Stefano Russo, Ege Cigirgan, Andrzej Chramiec-Głąbik, Łukasz Koziej, Cansu Cirzi, Jirka Peschek, Klaus Reuter, Mark Helm, Sebastian Glatt and Francesca Tuorto ()
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Wei Guo: Heidelberg University
Igor Kaczmarczyk: Jagiellonian University
Kevin Kopietz: Johannes Gutenberg-University Mainz
Florian Flegler: Philipps-Universität Marburg
Stefano Russo: Heidelberg University
Ege Cigirgan: Heidelberg University
Andrzej Chramiec-Głąbik: Jagiellonian University
Łukasz Koziej: Jagiellonian University
Cansu Cirzi: German Cancer Research Center (DKFZ)
Jirka Peschek: Heidelberg University
Klaus Reuter: Philipps-Universität Marburg
Mark Helm: Johannes Gutenberg-University Mainz
Sebastian Glatt: Jagiellonian University
Francesca Tuorto: DKFZ-ZMBH Alliance

Nature Communications, 2025, vol. 16, issue 1, 1-12

Abstract: Abstract 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.

Date: 2025
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DOI: 10.1038/s41467-025-62220-z

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