phaidra.vetmeduni.ac.at o:4906 DOI 10.1038/s44318-025-00443-y Determining the effects of pseudouridine incorporation on human tRNAs 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. eng tRNA Cryo-EM RNA Modifications RNA Folding Molecular Dynamics Simulation 2026-02-09T10:07:42.192Z LOMv1.0 Author BEGIN:VCARD VERSION:3.0 N:Biela;Anna; FN:Anna Biela END:VCARD BEGIN:VCARD VERSION:3.0 N:Nowak;Jakub; FN:Jakub Nowak END:VCARD BEGIN:VCARD VERSION:3.0 N:Biela;Artur; FN:Artur Biela END:VCARD BEGIN:VCARD VERSION:3.0 N:Mukherjee;Sunandan; FN:Sunandan Mukherjee END:VCARD BEGIN:VCARD VERSION:3.0 N:Moafinejad;Seyed Naeim; FN:Seyed Naeim Moafinejad END:VCARD BEGIN:VCARD VERSION:3.0 N:Maiti;Satyabrata; FN:Satyabrata Maiti END:VCARD BEGIN:VCARD VERSION:3.0 N:Chramiec-Glabik;Andrzej; FN:Andrzej Chramiec-Glabik END:VCARD BEGIN:VCARD VERSION:3.0 N:Mehta;Rahul; FN:Rahul Mehta END:VCARD BEGIN:VCARD VERSION:3.0 N:Jezowski;Jakub; FN:Jakub Jezowski END:VCARD BEGIN:VCARD VERSION:3.0 N:Dobosz;Dominika; FN:Dominika Dobosz END:VCARD BEGIN:VCARD VERSION:3.0 N:Dahate;Priyanka; FN:Priyanka Dahate END:VCARD BEGIN:VCARD VERSION:3.0 N:Arluison;Veronique; FN:Veronique Arluison END:VCARD BEGIN:VCARD VERSION:3.0 N:Wien;Frank; FN:Frank Wien END:VCARD BEGIN:VCARD VERSION:3.0 N:Indyka;Paulina; FN:Paulina Indyka END:VCARD BEGIN:VCARD VERSION:3.0 N:Rawski;Michal; FN:Michal Rawski END:VCARD BEGIN:VCARD VERSION:3.0 N:Bujnicki;Janusz M.; FN:Janusz M. Bujnicki END:VCARD BEGIN:VCARD VERSION:3.0 N:Lin;Ting-Yu; FN:Ting-Yu Lin END:VCARD BEGIN:VCARD VERSION:3.0 N:Glatt;Sebastian; FN:Sebastian Glatt X-ORCID:https://orcid.org/0000-0003-2815-7133 END:VCARD https://w3id.org/kim/hcrt/scheme https://w3id.org/kim/hcrt/text Textdokument Text LOMv1.0 yes https://creativecommons.org/licenses/by/4.0 application/pdf 10698157 https://phaidra.vetmeduni.ac.at/o:4906 https://phaidra.vetmeduni.ac.at/api/object/o:4906/thumbnail LOMv1.0 discipline