Comprehensive post-transcriptional modification profiles in individual Staphylococcus aureus tRNA species
Year: 2026
Type: article
Abstract: Post-transcriptional modifications modulate transfer RNA (tRNA) structure, stability, and codon decoding properties, contributing to translation regulation and adaptation across diverse organisms, including bacterial pathogens. We provide a comprehensive analysis of tRNA modifications in Staphylococcus aureus using extensive oligonucleotide mass spectrometry and deep-sequencing methods, generating a high-confidence modification map for each individual tRNA species, including non-proteogenic tRNAGly. While the overall tRNA modification landscape is conserved among Gram-positive bacteria, our data uncovered unexpected S. aureus-specific features. These include the absence of m2A37 in tRNAs despite the presence of the methyltransferase RlmN, a single multi-site DusB2 enzyme catalyzing all tRNA dihydrouridylation, and evidence suggesting a dedicated pseudouridine synthase responsible for Ψ32. Besides, heterogeneous modification patterns were observed in tRNALeu(UAA) and tRNALys(UUU), highlighting a complex interplay in anticodon hypermodification. Time-course proteomics revealed dynamic expression of tRNA modifying enzymes during growth. Integration of ribosome profiling and Nanopore tRNA sequencing offered a global view of S. aureus decoding properties, revealing efficient four-way wobble recognition, slower translation of rare codons by low abundant tRNAs, and distinctive decoding dynamics of Gly codons potentially influenced by the unusual modification status of tRNAGly(UCC). This work establishes a framework to dissect the role of tRNA modifications in S. aureus physiology and pathogenesis.
Source: Nucleic Acids Research
DOI: 10.1093/nar/gkag757
Authors: Jose R Jaramillo-Ponce, Philippe Wolff, Virginie Marchand, Yuri Motorin, Maximilian Kohl, Béatrice Chane-Woon-Ming, Hiroki Kanazawa, Aila Ruiz-Paterson, Caroline Paulus, Johana Chicher, Marine Lenon, Martina Krämer, Anne-Sophie Gribling-Burrer, Constantinos Stathopoulos, Redmond Smyth, Mark Helm, Pascale Romby, Stefano Marzi
Institutions: Centre National de la Recherche Scientifique, Architecture et Réactivité de l'arN, Université de Strasbourg, Ingénierie Moléculaire, Cellulaire et Physiopathologie, Université de Lorraine, Institut de Biologie Moléculaire et Cellulaire, Johannes Gutenberg University Mainz, University of Patras
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,992
Awards: G1094538864, G1474842019, G1512071772, G164444715, G1800105685, G2173752762, Par-delà les frontières, l'Université de Strasbourg, G2255650664, G2659019386, G2694206344, Façonner les talents en formation et en recherche à l'Université de Strasbourg, G2911418098, G2980570810, G3357951378, Structural analysis of Staphylococcus aureus ribonucleoprotein complex FemXAB-NPtRNAsGly essential for peptidoglycan synthesis and contributing to methicillin resistance, G3730343, G373712744, G399734812, G4381811279, Integrative Molecular and Cellular Biology, G4632815087, G471331628, G4779185514, G4964816082, TRR 319: RMaP: RNA Modifikation und Prozessierung, Αποκρυπτογράφηση της βιολογίας και των δυναμικών δικτύων νέων μη- κωδικοποιητικών RNAs στην κυτταρική ομοιοστασία και την αντίσταση σε αντικαρκινικά φάρμακα, Insectarium pour l'Infectiologie Moléculaire et Cellulaire, G6136357505, G6276897512, An integrated investigation of regulatory RNAs and their interacting proteins in bacteria models, G6803667113, G7240651115, RNA modifications in Staphylococcus aureus during stress responses, antibiotic treatments and virulence: impact on translation and its regulation, G8204089044, G8718739417
OpenAlex ID: W7197032826