Abstract
Vibrio cholerae is the causative agent of cholera, a dehydrating diarrheal disease. This Gram-negative pathogen is able to modulate its gene expression in order to combat stresses encountered in both aquatic and host environments, including stress posed by reactive oxygen species (ROS). In order to further the understanding of V. cholerae’s transcriptional response to ROS, we performed an RNA sequencing analysis to determine the transcriptional profile of V. cholerae when exposed to hydrogen hydroperoxide. Of 135 differentially expressed genes, VC0139 was amongst the genes with the largest induction. VC0139 encodes a protein homologous to the DPS (DNA-binding protein from starved cells) protein family, which are widely conserved and are implicated in ROS resistance in other bacteria. Using a promoter reporter assay, we show that during exponential growth, dps is induced by H2O2 in a manner dependent on the ROS-sensing transcriptional regulator, OxyR. Upon entry into stationary phase, the major stationary phase regulator RpoS is required to transcribe dps. Deletion of dps impaired V. cholerae resistance to both inorganic and organic hydroperoxides. Furthermore, we show that Dps is involved in resistance to multiple environmental stresses. Finally, we found that Dps is important for V. cholerae adult mouse colonization, but becomes dispensable in the presence of antioxidants. Taken together, our results suggest that Dps plays vital roles in both V. cholerae stress resistance and pathogenesis.
Highlights
The human pathogen Vibrio cholerae, a motile gram-negative bacterium, is the causative agent of the waterborne disease, cholera [1, 2] that is still a major threat to public health in developingPLOS ONE | DOI:10.1371/journal.pone.0171201 February 2, 2017Dps-mediated reactive oxygen species (ROS) resistance, and NIH/NIAID R01AI120489
In this study, using RNA sequencing and transcriptional reporters, we found that V. cholerae dps expression is induced by hydrogen peroxide in an OxyR-dependent manner
In order to study how V. cholerae manipulate their genetic reservoirs to resist oxidative stress, we analyzed the transcriptome of V. cholerae grown in the presence and absence of hydrogen
Summary
The human pathogen Vibrio cholerae, a motile gram-negative bacterium, is the causative agent of the waterborne disease, cholera [1, 2] that is still a major threat to public health in developingPLOS ONE | DOI:10.1371/journal.pone.0171201 February 2, 2017Dps-mediated ROS resistance (to B.K.), and NIH/NIAID R01AI120489 (to J.Z.). In this study, using RNA sequencing and transcriptional reporters, we found that V. cholerae dps expression is induced by hydrogen peroxide in an OxyR-dependent manner. For high osmolality stress assay, different growth-staged cultures of wild type and dps mutant cells were incubated in 1 M NaCl at 37 ̊C for 24 hrs.
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