The CpxRA two‐component system contributes to Legionella pneumophila virulence

Summary The bacterium Legionella pneumophila is capable of intracellular replication within freshwater protozoa as well as human macrophages, the latter of which results in the serious pneumonia Legionnaires’ disease. A primary factor involved in these host cell interactions is the Dot/Icm Type IV secretion system responsible for translocating effector proteins needed to establish and maintain the bacterial replicative niche. Several regulatory factors have been identified to control the expression of the Dot/Icm system and effectors, one of which is the CpxRA two‐component system, suggesting essentiality for virulence. In this study, we generated cpxR, cpxA and cpxRA in‐frame null mutant strains to further delineate the role of the CpxRA system in bacterial survival and virulence. We found that cpxR is essential for intracellular replication within Acanthamoeba castellanii, but not in U937‐derived macrophages. Transcriptome analysis revealed that CpxRA regulates a large number of virulence‐associated proteins including Dot/Icm effectors as well as Type II secreted substrates. Furthermore, the cpxR and cpxRA mutant strains were more sodium resistant than the parental strain Lp02, and cpxRA expression reaches maximal levels during postexponential phase. Taken together, our findings suggest the CpxRA system is a key contributor to L. pneumophila virulence in protozoa via virulence factor regulation. Overview of the CpxRA regulon and its role in L. pneumophila virulence. We identified the CpxRA system to be essential for survival in protozoa, but not in human macrophages. Expression of cpxRA, genetically organized in an operon structure with three other genes lpg1439‐1441, is subjected to positive autoregulation and negative regulation by an unknown factor. Transcriptomic analyses expanded the list of CpxRA regulon members to include Type II secretion system substrates and additional Dot/Icm effectors.