The RNA chaperone Hfq is a novel regulator of catalase expression and hydrogen peroxide-induced oxidative stress response in Listeria monocytogenes EGD-e

Abstract

The RNA chaperone Hfq plays a pivotal role in many bacteria, acting as a regulator of gene expression and promoting interaction between mRNA-sRNA pairs in Gram-negative bacteria. However, in Gram-positive bacteria this protein is expendable for riboregulation, and the main function of Hfq remains elusive. This work unveils a novel function for Hfq in the oxidative stress response of the human pathogen Listeria monocytogenes, a Gram-positive bacterium responsible for the infectious disease listeriosis. Disruption of hfq gene (Δhfq) results in a hypersensitive phenotype towards hydrogen peroxide (H₂O₂), in which sub-inhibitory concentrations of this reactive oxygen species (ROS) severely impair growth and viability of L. monocytogenes EGD-e. A Δhfq-complemented strain does not show this phenotype. This Hfq-dependent regulation of oxidative stress seems specific for H₂O₂, as exposure to superoxides caused no differences. We demonstrate that Hfq has a dual regulatory role in the expression of catalase (kat), the key enzyme involved in H₂O₂ detoxification. Hfq influences kat transcription under non-stress conditions by modulating the levels of the transcriptional repressor PerR, and also acts post-transcriptionally by stabilizing kat mRNA under H₂O₂-induced stress. Indeed, enzymatic assays revealed reduced catalase activity in Δhfq cell extracts, a result unrelated to differences in cellular iron content. Bacterial infection triggers immune cells to produce massive amounts of ROS, like H₂O₂. We show that inactivation of Hfq increases susceptibility to macrophage killing, connecting Hfq with the stress resistance and virulence of L. monocytogenes EGD-e. Overall, these findings advance the understanding of Hfq function within Gram-positive bacteria, revealing for the first time that Hfq is a novel regulator of catalase expression. This paves the way for the study of yet unknown oxidative stress response pathways regulated by Hfq in other pathogens.

Keywords: Bacterial virulence; Catalase; Hfq; Hydrogen peroxide; Oxidative stress; PerR.