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Brucella abortus senses the intracellular environment through the two-component system BvrR/BvrS allowing the adaptation to its replicative niche.

Infection and Immunity 2018 January 30
Brucella abortus is a facultative extracellular-intracellular pathogen belonging a group of α-Proteobacteria that establishes close interactions with animal cells. This bacterium enters host cells in a membrane bound compartment, avoiding the lysosomal route and reaching the endoplasmic reticulum through the action of the Type IV secretion system, VirB. In this work we demonstrate that the two-component system BvrR/BvrS senses the intracellular environment to mount the transcriptional response required for intracellular life adaptation. By combining a method to purify intracellularly extracted bacteria with a strategy that allows direct determination of BvrR phosphorylation we showed that upon entrance to host cells, the regulatory protein BvrR was activated (BvrR-P) by phosphorylation at aspartate 58. This activation takes place in response to intracellular cues found in early compartments, such as low pH and nutrient deprivation. Furthermore, BvrR activation was followed by an increase in the expression of VjbR and VirB. The in vitro activation of this BvrR-P/VjbR/VirB virulence circuit rescued B. abortus from the inhibition of intracellular replication induced by bafilomycin treatment of cells, demonstrating the relevance of this mechanism for the intracellular bacterial survival and replication. Altogether, our results indicate that B. abortus senses the transition from the extracellular to the intracellular milieu through BvrR/BvrS allowing the bacterium to transit safely to its replicative niche. These results serve as a working model for understanding the role of this family of two-component systems in the adaptation to intracellular life of α-Proteobacteria.

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