The BvgS PAS domain: an independent sensory perception module in theBordetellaBvgAS phosphorelay

To detect and respond to the diverse environments they encounter, bacteria often use two-component regulatory systems (TCSs) to coordinate essential cellular processes required for survival. In pathogenic Bordetella species, the BvgAS TCS regulates expression of hundreds of genes, including those encoding all known protein virulence factors, and its kinase activity is essential for respiratory infection. Maintenance of BvgS kinase activity in the lower respiratory tract (LRT) depends on the function of another TCS, PlrSR. While the periplasmic venus fly-trap domains of BvgS have been implicated in responding to so-called modulating signals in vitro (nicotinic acid and MgSO 4 ), a role for the cytoplasmic Per-Arnt-Sim (PAS) domain in signal perception has not previously been demonstrated. By comparing B. bronchiseptica strains with mutations in the PAS domain-encoding region of bvgS with wild-type bacteria in vitro and in vivo , we found that although the PAS domain is not required to sense modulating signals in vitro , it is required for the inactivation of BvgS that occurs in the absence of PlrS in the LRT of mice, suggesting that the BvgS PAS domain functions as an independent signal perception domain. Our data also indicate that the BvgS PAS domain is important for controlling absolute levels of BvgS kinase activity and the efficiency of the response to modulating signals in vitro . Our results indicate that BvgS is capable of integrating sensory inputs from both the periplasm and the cytoplasm to control precise gene expression patterns in diverse environmental conditions. Importance Despite high rates of vaccination, Pertussis, a severe, highly contagious respiratory disease, caused by the bacterium Bordetella pertussis , has reemerged as a significant health threat. In Bordetella pertussis and the closely related species, Bordetella bronchiseptica , activity of the BvgAS two-component regulatory system is critical for colonization of the human respiratory tract and other mammalian hosts, respectively. Here we show that the cytoplasmic PAS domain of BvgS can function as an independent signal perception domain that is capable of integrating environmental signals that influence overall BvgS activity. Our work is significant as it reveals a critical, yet previously unrecognized role, for the PAS domain in the BvgAS phosphorelay and provides a greater understanding of virulence regulation in Bordetella .