Comprehensive post-transcriptional modification profiles in individual Staphylococcus aureus tRNA species
Abstract
Post-transcriptional modifications modulate transfer RNA (tRNA) structure, st abilit y, 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 tRNA Gly . 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 m 2 A37 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 32. Besides, heterogeneous modification patterns were observed in tRNA Leu (UAA) and tRNA Lys (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 tRNA Gly (UCC). This work establishes a framework to dissect the role of tRNA modifications in S. aureus physiology and pathogenesis.
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BibTeX
@article{jaramilloponce2026comprehensive,
title = {{Comprehensive post-transcriptional modification profiles in individual Staphylococcus aureus tRNA species}},
author = {Jose R Jaramillo-Ponce and Philippe Wolff and Virginie Marchand and Yuri Motorin and Maximilian Kohl and Béatrice Chane-Woon-Ming and Hiroki Kanazawa and Aila Ruiz-Paterson and Caroline Alexandra Paulus and Johana Chicher and Marine Lénon and Martina Krämer and Anne-Sophie Gribling-Burrer and Constantinos Stathopoulos and Redmond Smyth and Mark Helm and Pascale Romby and Stefano Marzi},
journal = {Nucleic Acids Research},
year = {2026},
month = aug,
doi = {10.1093/nar/gkag757},
url = {https://univoak.hal.science/hal-05758373v1},
note = {CC BY-NC 4.0},
}
RIS
TY - JOUR
TI - Comprehensive post-transcriptional modification profiles in individual Staphylococcus aureus tRNA species
AU - Jose R Jaramillo-Ponce
AU - Philippe Wolff
AU - Virginie Marchand
AU - Yuri Motorin
AU - Maximilian Kohl
AU - Béatrice Chane-Woon-Ming
AU - Hiroki Kanazawa
AU - Aila Ruiz-Paterson
AU - Caroline Alexandra Paulus
AU - Johana Chicher
AU - Marine Lénon
AU - Martina Krämer
AU - Anne-Sophie Gribling-Burrer
AU - Constantinos Stathopoulos
AU - Redmond Smyth
AU - Mark Helm
AU - Pascale Romby
AU - Stefano Marzi
PY - 2026
DA - 2026/08/06
JO - Nucleic Acids Research
DO - 10.1093/nar/gkag757
UR - https://univoak.hal.science/hal-05758373v1
AB - Post-transcriptional modifications modulate transfer RNA (tRNA) structure, st abilit y, 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 tRNA Gly . 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 m 2 A37 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 32. Besides, heterogeneous modification patterns were observed in tRNA Leu (UAA) and tRNA Lys (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 tRNA Gly (UCC). This work establishes a framework to dissect the role of tRNA modifications in S. aureus physiology and pathogenesis.
SN - 0305-1048
SN - 1362-4962
ER -