The Butyrivibrio fibrisolvens tet(W) gene is located on the conjugative transposon TnB1230. TnB1230 encodes transfer proteins with 48 to 67% identity to some of those encoded by Tn1549. tet(W) is flanked by directly repeated sequences with significant homology to oxygen-insensitive nitroreductases. The 340 nucleotides upstream of tet(W) are strongly conserved and are required for tetracycline resistance.Tetracyclines are the second most widely used group of antibiotics worldwide, and tetracycline resistance (Tc r ) is extremely common among bacteria (17). Many Tc r genes are transmissible, being carried by plasmids or transposable chromosomal elements. Tc r genes have mainly been described for pathogenic bacteria, but the new resistance genes tet(W) (2, 20) and tet(32) (12) were first identified in anaerobic commensal bacteria from the rumen and from porcine and human feces. These genes are distinct from previously described ribosome protection genes, with the highest identities being 68% [to Tet(W)] and 76% [to Tet(32)] at the corresponding amino acid level. tet(W) is one of the most widespread tetracycline resistance genes in environmental samples (1,23).Copies of tet(W) found in environmentally and phylogenetically distinct bacterial isolates (2, 20) have remarkable sequence conservation, which argues for the extensive rapid transfer of this gene in nature. Previous work showed that tet(W) could be transferred at high frequencies (10 Ϫ2 to 10 Ϫ5

Trans-translation is a ubiquitous bacterial mechanism for ribosome rescue in the event of translation stalling. Although trans-translation is not essential in several bacterial species, it has been found essential for viability or virulence in a wide range of pathogens. We describe here that trans-translation is essential in the human pathogen Legionella pneumophila, the etiologic agent of Legionnaire’s disease (LD), a severe form of nosocomial and community-acquired pneumonia. The ssrA gene coding for tmRNA, the key component of trans-translation, could not be deleted in L. pneumophila. To circumvent this and analyse the consequences of impaired trans-translation, we placed ssrA under the control of a chemical inducer. Phenotypes associated with the inhibition of ssrA expression include growth arrest in rich medium, hampered cell division, and hindered ability to infect eukaryotic cells (amoebae and human macrophages). LD is often associated with failure of antibiotic treatment and death (>10% of clinical cases). Decreasing tmRNA levels led to significantly higher sensitivity to ribosome-targeting antibiotics, including to erythromycin. We also detected a higher sensitivity to the transcription inhibitor rifampicin. Both antibiotics are recommended treatments for LD. Thus, interfering with trans-translation may not only halt the infection, but could also potentiate the recommended therapeutic treatments of LD.