The molecular basis of tRNA selectivity by human pseudouridine synthase 3.

Autor:	Lin TY; Małopolska Centre of Biotechnology, Jagiellonian University, 30-387 Kraków, Poland. Electronic address: ting-yu.lin@durham.ac.uk., Kleemann L; Department of Chemistry, Biochemistry and Pharmaceutical Sciences, University of Bern, 3012 Bern, Switzerland; Graduate School for Cellular and Biomedical Sciences, University of Bern, 3012 Bern, Switzerland., Jeżowski J; Małopolska Centre of Biotechnology, Jagiellonian University, 30-387 Kraków, Poland; Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Kraków, Poland., Dobosz D; Małopolska Centre of Biotechnology, Jagiellonian University, 30-387 Kraków, Poland., Rawski M; Małopolska Centre of Biotechnology, Jagiellonian University, 30-387 Kraków, Poland; SOLARIS National Synchrotron Radiation Centre, Jagiellonian University, 30-392 Kraków, Poland., Indyka P; Małopolska Centre of Biotechnology, Jagiellonian University, 30-387 Kraków, Poland; SOLARIS National Synchrotron Radiation Centre, Jagiellonian University, 30-392 Kraków, Poland., Ważny G; SOLARIS National Synchrotron Radiation Centre, Jagiellonian University, 30-392 Kraków, Poland; Doctoral School of Exact and Natural Sciences, Jagiellonian University, 30-348 Kraków, Poland., Mehta R; Małopolska Centre of Biotechnology, Jagiellonian University, 30-387 Kraków, Poland; Doctoral School of Exact and Natural Sciences, Jagiellonian University, 30-348 Kraków, Poland., Chramiec-Głąbik A; Małopolska Centre of Biotechnology, Jagiellonian University, 30-387 Kraków, Poland., Koziej Ł; Małopolska Centre of Biotechnology, Jagiellonian University, 30-387 Kraków, Poland., Ranff T; Institute of Pharmacy and Molecular Biotechnology, Heidelberg University, 69120 Heidelberg, Germany., Fufezan C; Institute of Pharmacy and Molecular Biotechnology, Heidelberg University, 69120 Heidelberg, Germany., Wawro M; Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Kraków, Poland., Kochan J; Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Kraków, Poland., Bereta J; Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Kraków, Poland., Leidel SA; Department of Chemistry, Biochemistry and Pharmaceutical Sciences, University of Bern, 3012 Bern, Switzerland; Graduate School for Cellular and Biomedical Sciences, University of Bern, 3012 Bern, Switzerland. Electronic address: sebastian.leidel@unibe.ch., Glatt S; Małopolska Centre of Biotechnology, Jagiellonian University, 30-387 Kraków, Poland. Electronic address: sebastian.glatt@uj.edu.pl.
Jazyk:	angličtina
Zdroj:	Molecular cell [Mol Cell] 2024 Jul 11; Vol. 84 (13), pp. 2472-2489.e8.
DOI:	10.1016/j.molcel.2024.06.013
Abstrakt:	Pseudouridine (Ψ), the isomer of uridine, is ubiquitously found in RNA, including tRNA, rRNA, and mRNA. Human pseudouridine synthase 3 (PUS3) catalyzes pseudouridylation of position 38/39 in tRNAs. However, the molecular mechanisms by which it recognizes its RNA targets and achieves site specificity remain elusive. Here, we determine single-particle cryo-EM structures of PUS3 in its apo form and bound to three tRNAs, showing how the symmetric PUS3 homodimer recognizes tRNAs and positions the target uridine next to its active site. Structure-guided and patient-derived mutations validate our structural findings in complementary biochemical assays. Furthermore, we deleted PUS1 and PUS3 in HEK293 cells and mapped transcriptome-wide Ψ sites by Pseudo-seq. Although PUS1-dependent sites were detectable in tRNA and mRNA, we found no evidence that human PUS3 modifies mRNAs. Our work provides the molecular basis for PUS3-mediated tRNA modification in humans and explains how its tRNA modification activity is linked to intellectual disabilities. Competing Interests: Declaration of interests The authors declare no competing interests. (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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