Quorum sensing LuxR proteins VjbR and BabR jointly regulate Brucella abortus survival during infection

Brucella abortus maintains an N-acetyl homoserine lactone quorum sensing system that consists of two LuxR proteins, VjbR and BabR, as well as two signals, dodecanoyl (C12 AHL) and 3-oxododecanoyl (3-OXO-C12 AHL) homoserine lactone. This system regulates major virulence factors that influence the bacteria's survival during infection. We generated the first strain that lacks both LuxR proteins and found a synergistic interaction for survival in the chronic infection C57BL/6 mouse model. Transcriptomic analyses of the ∆vjbR∆babR double-deletion strain, as well as the cognate single-deletion strains, in a rich medium with vehicle control or supplemented with an AHL signal revealed large-scale genetic dysregulation in all conditions. Moreover, the double mutant maintained a limited response to AHL, even in the absence of the LuxR proteins. We additionally found that quorum sensing regulates the denitrification pathway but found no in vitro differences in the ability of quorum sensing deletion strains to clear nitric oxide stress or grow under anoxic denitrifying conditions. Finally, we confirmed that BabR autoregulates its own expression, and that VjbR mildly represses BabR expression. Altogether, these experiments help further characterize the Brucella quorum sensing systems and indicate that further attention should be given to the joint interactions between VjbR and BabR in controlling virulence.IMPORTANCEBrucella abortus is a zoonotic bacterial pathogen that uses its quorum sensing to survive within hosts. This study further characterizes that system and indicates important future lines of inquiry. We found that both quorum sensing proteins, VjbR and BabR, coordinate to maintain survival, as well as document that both quorum sensing systems appear physiologically active.