Determining the effects of pseudouridine incorporation on human tRNAs

Biela, Anna D; Nowak, Jakub S; Biela, Artur P; Mukherjee, Sunandan; Moafinejad, Seyed Naeim; Maiti, Satyabrata; Chramiec-Głąbik, Andrzej; Mehta, Rahul; Jeżowski, Jakub; Dobosz, Dominika; Dahate, Priyanka; Arluison, Veronique; Wien, Frank; Indyka, Paulina; Rawski, Michal; Bujnicki, Janusz M; Lin, Ting-Yu; Glatt, Sebastian

Abstract

Transfer RNAs (tRNAs) are ubiquitous non-coding RNA molecules required to translate mRNA-encoded sequence information into nascent polypeptide chains. Their relatively small size and heterogenous patterns of their RNA modifications have impeded the systematic structural characterization of individual tRNAs. Here, we use single-particle cryo-EM to determine the structures of four human tRNAs before and after incorporation of pseudouridines (Ψ). Following post-transcriptional modifications by distinct combinations of human pseudouridine synthases, we find that tRNAs become stabilized and undergo specific local structural changes. We establish interactions between the D- and T-arms as the key linchpin in the tertiary structure of tRNAs. Our structures of human tRNAs highlight the vast potential of cryo-EM combined with biophysical measurements and computational simulations for structure-function analyses of tRNAs and other small, folded RNA domains.

Citation

Biela, A. D., Nowak, J. S., Biela, A. P., Mukherjee, S., Moafinejad, S. N., Maiti, S., Chramiec-Głąbik, A., Mehta, R., Jeżowski, J., Dobosz, D., Dahate, P., Arluison, V., Wien, F., Indyka, P., Rawski, M., Bujnicki, J. M., Lin, T.-Y., & Glatt, S. (online). Determining the effects of pseudouridine incorporation on human tRNAs. The EMBO Journal, https://doi.org/10.1038/s44318-025-00443-y

• Journal Article Type: Article
• Acceptance Date: Apr 2, 2025
• Online Publication Date: Apr 29, 2025
• Deposit Date: Apr 30, 2025
• Publicly Available Date: May 14, 2025
• Journal: The EMBO Journal
• Print ISSN: 0261-4189
• Electronic ISSN: 1460-2075
• Publisher: EMBO Press
• Peer Reviewed: Peer Reviewed
• DOI: https://doi.org/10.1038/s44318-025-00443-y
• Public URL: https://durham-repository.worktribe.com/output/3806520