The FUR-like regulators PerRA and PerRB integrate a complex regulatory network that promotes mammalian host-adaptation and virulence of Leptospira interrogans.

André A Grassmann,Melissa J Caimano,Everton B Bettin,Crispin Zavala-Alvarado,Mathieu Picardeau,Nadia Benaroudj

doi:10.1371/journal.ppat.1009078

Abstract

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 compared L. interrogans cultivated in vitro and in vivo using 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 in Bacillus 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. Using perRA and perRB single and double mutants, we establish that L. interrogans requires at least one functional PerR for infectivity and renal colonization in a reservoir host. Our finding that the perRA/B double 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 of perRA, perRB and perRA/B mutants cultivated within DMCs identified 106 genes that are dysregulated in the double mutant, including ligA, ligB and lvrA/B sensory histidine kinases. Decreased expression of LigA and LigB in the perRA/B mutant was not due to loss of LvrAB signaling. The majority of genes in the perRA and perRB single and double mutant DMC regulons were differentially expressed only in vivo, highlighting the importance of host signals for regulating gene expression in L. 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 by L. interrogans within mammals.

Highlights

Leptospirosis is a neglected tropical disease with a worldwide distribution [1, 2]
The majority of cases of human leptospirosis are associated with Leptospira interrogans
We identified >100 genes that were dysregulated in the perRB Tn mutant; the resulting double mutant (perRA/B) double mutant cultivated in rat peritoneal cavities, including the virulence determinants LigA and LigB

Summary

Introduction

Leptospirosis is a neglected tropical disease with a worldwide distribution [1, 2]. Globally, ~1 million cases and ~60,000 deaths are estimated each year [3]. The majority of severe cases of human leptospirosis are associated with L. interrogans [9]. Infected reservoir hosts shed large numbers of leptospires (up to 107/mL) in their urine for weeks to months [9, 12, 14, 15]. While infection in reservoir hosts is generally asymptomatic, incidental hosts, including humans, may develop clinical symptoms ranging from mild flu-like illness to fulminant disease (e.g., Weil’s disease and pulmonary hemorrhage syndrome). The factors driving disease severity in humans are poorly understood but are thought to include the bacterial serovar and strain, inoculum size, and the host’s innate and adaptive immune responses [6, 16,17,18]

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Conclusion