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.
Abstract
Journal Article
eng
38996458
Lin, Ting-Yu, et al. "The Molecular Basis of tRNA Selectivity By Human Pseudouridine Synthase 3." Molecular Cell, vol. 84, no. 13, 2024, pp. 2472-2489.e8.
Lin TY, Kleemann L, Jeżowski J, et al. The molecular basis of tRNA selectivity by human pseudouridine synthase 3. Mol Cell. 2024;84(13):2472-2489.e8.
Lin, T. Y., Kleemann, L., Jeżowski, J., Dobosz, D., Rawski, M., Indyka, P., Ważny, G., Mehta, R., Chramiec-Głąbik, A., Koziej, Ł., Ranff, T., Fufezan, C., Wawro, M., Kochan, J., Bereta, J., Leidel, S. A., & Glatt, S. (2024). The molecular basis of tRNA selectivity by human pseudouridine synthase 3. Molecular Cell, 84(13), 2472-e8. https://doi.org/10.1016/j.molcel.2024.06.013
Lin TY, et al. The Molecular Basis of tRNA Selectivity By Human Pseudouridine Synthase 3. Mol Cell. 2024 Jul 11;84(13):2472-2489.e8. PubMed PMID: 38996458.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR
T1 - The molecular basis of tRNA selectivity by human pseudouridine synthase 3.
AU - Lin,Ting-Yu,
AU - Kleemann,Leon,
AU - Jeżowski,Jakub,
AU - Dobosz,Dominika,
AU - Rawski,Michał,
AU - Indyka,Paulina,
AU - Ważny,Grzegorz,
AU - Mehta,Rahul,
AU - Chramiec-Głąbik,Andrzej,
AU - Koziej,Łukasz,
AU - Ranff,Tristan,
AU - Fufezan,Christian,
AU - Wawro,Mateusz,
AU - Kochan,Jakub,
AU - Bereta,Joanna,
AU - Leidel,Sebastian A,
AU - Glatt,Sebastian,
PY - 2023/04/25/received
PY - 2024/03/14/revised
PY - 2024/06/13/accepted
PY - 2024/7/13/medline
PY - 2024/7/13/pubmed
PY - 2024/7/12/entrez
KW - PUS1
KW - PUS3
KW - Pseudo-seq
KW - anticodon stem loop
KW - cryo-EM structure
KW - pseudouridine synthase
KW - tRNA modification
KW - transcriptome
SP - 2472
EP - 2489.e8
JF - Molecular cell
JO - Mol Cell
VL - 84
IS - 13
N2 - 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.
SN - 1097-4164
UR - https://www.unboundmedicine.com/prime/citation/38996458/The_molecular_basis_of_tRNA_selectivity_by_human_pseudouridine_synthase_3.
DB - PRIME
DP - Unbound Medicine
ER -
