phaidra.vetmeduni.ac.at o:4844 DOI 10.1080/15476286.2025.2541421 Mechanistic insight into the pseudouridylation of RNA Pseudouridylation (Ψ) is a highly abundant and conserved RNA modification that is present in all three domains of life. The incorporation of Ψ can affect the stability of RNAs, modulate their interaction patterns, and regulate many other aspects of RNA biology. Ψ are introduced by a structurally highly related enzyme family, called pseudouridine synthases (PUS). Each PUS enzyme targets distinct RNA substrates and target sites. Dysregulation of PUS enzymes has been implicated in neurodevelopmental disorders, mitochondrial diseases and cancer. These clinical consequences highlight the ultimate need to understand how these enzymes catalyze their modification reactions and achieve target selectivity as well as specificity. In this review, we summarize the currently available structural information on PUS enzymes and highlight the most recent progress in understanding some underlying mechanistic principles. Furthermore, we illustrate the increasing therapeutic applications related to the so-called 5th RNA base. eng Pseudouridine tRNA RNA Modification Pseudouridine Synthase Cryo-electron Microscopy Crystallography Neurodegenerative Disease Cancer 2026-02-02T15:38:40.920Z LOMv1.0 Author BEGIN:VCARD VERSION:3.0 N:Lin;Ting-Yu; FN:Ting-Yu Lin END:VCARD BEGIN:VCARD VERSION:3.0 N:Stone;Yasmin; FN:Yasmin Stone 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-nc/4.0 application/pdf 4651319 https://phaidra.vetmeduni.ac.at/o:4844 https://phaidra.vetmeduni.ac.at/api/object/o:4844/thumbnail LOMv1.0 discipline