Loss of RNA Chaperone Hfq Unveils a Toxic Pathway in Pseudomonas aeruginosa.

Hfq is an RNA chaperone that serves as a master regulator of bacterial physiology. Here we show that in the opportunistic pathogen , the loss of Hfq can result in a dramatic reduction in growth in a manner that is dependent upon MexT, a transcription regulator that governs antibiotic resistance in this organism. Using a combination of chromatin immunoprecipitation with high-throughput sequencing and transposon insertion sequencing, we identify the MexT-activated genes responsible for mediating the growth defect of mutant cells. These include a newly identified MexT-controlled gene that we call We demonstrate that encodes a small protein that is acutely toxic to wild-type cells when produced ectopically. Furthermore, we show that expression is negatively regulated by Hfq, offering a possible explanation for the growth defect of mutant cells. Finally, we present evidence that the expression of MexT-activated genes is dependent upon GshA, an enzyme involved in the synthesis of glutathione. Our findings suggest that Hfq can influence the growth of by limiting the toxic effects of specific MexT-regulated genes. Moreover, our results identify glutathione to be a factor important for the activity of MexT. Here we show that the conserved RNA chaperone Hfq is important for the growth of the opportunistic pathogen We found that the growth defect of mutant cells is dependent upon the expression of genes that are under the control of the transcription regulator MexT. These include a gene that we refer to as , which we show is negatively regulated by Hfq and encodes a small protein that can be toxic when ectopically produced in wild-type cells. Thus, Hfq can influence the growth of by limiting the toxic effects of MexT-regulated genes, including one encoding a previously unrecognized small protein. We also show that MexT activity depends on an enzyme that synthesizes glutathione.