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Loss of s2U tRNA modification induces antibiotic tolerance in Yersinia pseudotuberculosis

Cotten, K. L.; McShane, A.; Dedon, P. C.; Begley, T. J.; Davis, K. M.

2025-07-01 microbiology
10.1101/2025.07.01.662621 bioRxiv
Show abstract

Stress promotes phenotypic changes in bacteria that allow them to survive antibiotic treatment. This phenomenon, termed antibiotic tolerance, can cause treatment failure, highlighting a need to define bacterial pathways that promote survival. Previously, we found Yersinia pseudotuberculosis downregulates tusB, a gene involved in modifying tRNAs with s2U, in response to doxycycline. Here we find that deletion of tusB results in loss of s2U and induces antibiotic tolerance. Using a combination of sequencing-based approaches and analysis of gene codon usage, our data show that loss of s2U decreases translation of ribosomal proteins. Ribosomal proteins are highly enriched in codons that require s2U-modified tRNAs for efficient translation, and loss of s2U results in ribosome pausing at these codons. Our results highlight a previously unknown mechanism of antibiotic tolerance where reduction in ribosomal protein abundance can globally reduce translation, and describes a novel strategy bacteria use to slow growth by modulating s2U levels.

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