The FUR-like regulators PerRA and PerRB integrate a complex regulatory network that promotes mammalian host-adaptation and virulence ofLeptospira interrogans
Authors: Grassmann André A. , Zavala-Alvarado Crispin , Bettin Everton B. , Picardeau Mathieu , Benaroudj Nadia , Caimano Melissa J. 
Published: November 2020

Leptospira interrogans, the causative agent of most cases of human leptospirosis, must respond to myriad environmental signals during its free-living and pathogenic lifestyles. Previously, we comparedL. interroganscultivatedin vitroandin vivousing a dialysis membrane chamber (DMC) peritoneal implant model. From these studies emerged the importance of genes encoding the Peroxide responsive regulators PerRA and PerRB. First described in inBacillus subtilis, PerRs are widespread in Gram-negative and -positive bacteria, where regulate the expression of gene products involved in detoxification of reactive oxygen species and virulence. UsingperRAandperRBsingle and double mutants, we establish thatL. interrogansrequires at least one functional PerR for infectivity and renal colonization in a reservoir host. Our finding that theperRA/Bdouble mutant survives at wild-type levels in DMCs is noteworthy as it demonstrates that the loss of virulence is not due to a metabolic lesion (i.e., metal starvation) but instead reflects dysregulation of virulence-related gene products. Comparative RNA-Seq analyses ofperRA,perRBandperRA/Bmutants cultivated within DMCs identified 106 genes that are dysregulated in the double mutant, includingligA, ligBandlvrA/Bsensory histidine kinases. Decreased expression of LigA and LigB in theperRA/Bmutant was not due to loss of LvrAB signaling. The majority of genes in theperRAandperRBsingle and double mutant DMC regulons were differentially expressed onlyin vivo, highlighting the importance of host signals for regulating gene expression inL. interrogans. Importantly, the PerRA, PerRB and PerRA/B DMC regulons each contain multiple genes related to environmental sensing and/or transcriptional regulation. Collectively, our data suggest that PerRA and PerRB are part of a complex regulatory network that promotes host adaptation byL. interroganswithin mammals.

Author Summary

Leptospirosis is a neglected tropical disease with a worldwide distribution. Globally, ~1 million cases and ~60,000 deaths are reported each year. The majority of cases of human leptospirosis are associated withLeptospira interrogans. Infection begins when a naïve reservoir (or incidental) host comes into direct or indirect contact with urine from an infected reservoir host. While infection in reservoir hosts, including rats and mice, is generally asymptomatic, incidental hosts, including humans, may develop clinical symptoms ranging from mild flu-like illness to fulminant disease. The gene products required by leptospires for infection remain poorly understood. Herein, we establish that the FUR family regulators PerRA and PerRB function in parallel, contributing to infectivity and renal colonization in mice. By comparative transcriptomics, we identified >100 genes that were dysregulated in theperRA/Bdouble mutant cultivated in rat peritoneal cavities, including the virulence determinants LigA and LigB. Importantly, the PerRA, PerRB and PerRA/B DMC regulons contain multiple genes related to environmental sensing and/or transcriptional regulation. Our data suggest that PerRA and PerRB are part of a complex regulatory network that promotes host adaptation byL. interroganswithin mammals.