PXD060249

PXD060249 is an original dataset announced via ProteomeXchange.

Title	tRNA hydroxylation is an epitranscriptomic modulator of metabolic states affecting Pseudomonas aeruginosa pathogenicity
Description	Post-transcriptional modification of tRNAs represents an essential layer of translational regulation critical for bacterial adaptation to environmental changes. Increasing evidence links the tRNA epitranscriptome to pivotal roles in the regulation of gene expression and various cellular processes, including stress responses and the establishment of virulence phenotypes. In this study, we used mass-spectrometry and nanopore sequencing to systematically identify and quantify sites of TrhPO-dependent tRNA hydroxylation in total and purified tRNAs in the opportunistic human pathogen Pseudomonas aeruginosa. Furthermore, transcriptomic, ribosome profiling and proteomic data were integrated to demonstrate the post-transcriptional consequences of the absence of xo5U modifications at wobble positions in selected tRNAs. We show that changes in metabolic fluxes drive the impaired ability to infect host cells and strongly attenuated virulence in the Galleria mellonella model. In the absence of TrhPO-mediated tRNA modification, chorismate, the precursor for the biosynthesis of xo5U-modifications, is funneled into alternative pathways including the production of aromatic amino acids and phenazines. Our findings that metabolic rerouting rather than changes in proteome profiles attenuates P. aeruginosa virulence highlights the multifunctional roles of tRNA-modifying enzymes and suggests an underexplored role for these enzymes in monitoring and modulating metabolic fitness. These insights open new avenues for combatting the pathogenicity of this challenging opportunistic pathogen.
HostingRepository	PRIDE
AnnounceDate	2025-08-04
AnnouncementXML	Submission_2025-08-03_16:07:14.639.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Benedikt Kennepohl
SpeciesList	scientific name: Pseudomonas aeruginosa (strain UCBPP-PA14); NCBI TaxID: 208963;
ModificationList	acetylated residue; monohydroxylated residue; deamidated residue
Instrument	Orbitrap Exploris 240

Revision	Datetime	Status	ChangeLog Entry
0	2025-01-27 06:04:45	ID requested
⏵ 1	2025-08-03 16:07:15	announced

Frommeyer YN, Gomez NO, Preusse M, Arce-Rodriguez A, Neubauer K, Kennepohl B, Witte J, Bouheraoua S, Cetraro P, Erdmann J, Neumann-Schaal M, M, ü, sken M, Pich A, B, ä, hre H, Depledge DP, H, ä, ussler S, tRNA hydroxylation is an epitranscriptomic modulator of metabolic states affecting Pseudomonas aeruginosa pathogenicity. Nucleic Acids Res, 53(14):(2025) [pubmed]

10.1093/nar/gkaf719;

submitter keyword: tRNA modification,Pseudomonas aeruginosa, epitranscriptomics

Susanne Häußler
contact affiliation	Helmholtz Centre for Infection Research Dept. of Molecular Bacteriology Inhoffenstr. 7 D-38124 Braunschweig Germany
contact email	susanne.haeussler@helmholtz-hzi.de
lab head
Benedikt Kennepohl
contact affiliation	Twincore Hannover
contact email	benedikt.kennepohl@helmholtz-hzi.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD060249
     1. Label: PRIDE project
     2. Name: tRNA hydroxylation is an epitranscriptomic modulator of metabolic states affecting Pseudomonas aeruginosa pathogenicity