The Legionella Lqs-LvbR regulatory network controls temperature-dependent growth onset and bacterial cell density

Abstract

Legionella species are facultative intracellular pathogens, which can cause a life-threatening pneumonia termed Legionnaires disease. Legionella pneumophila employs the Legionella quorum sensing (Lqs)-LvbR network to regulate virulence and motility, but its role for growth in media is ill-defined. Compared to the parental L. pneumophila strain JR32, a {Delta}lqsR mutant showed a reduced lag phase at 30{degrees}C and reached a higher cell density at 45{degrees}C, while the {Delta}lqsA, {Delta}lqsS and {Delta}lqsT mutants exhibited a longer lag phase and reached only a lower cell density. A {Delta}lvbR mutant resumed growth like the parental strain at 30{degrees}C, but exhibited a substantially reduced cell density at 45{degrees}C. Thus, LvbR is an important cell density regulator at elevated temperatures. A quantitative analysis of temperature-dependent growth characteristics of environmental and clinical strains revealed that L. pneumophila strains grew in AYE medium after distinct lag phases with similar rates at 30{degrees}C, reached different cell densities at the optimal growth temperature of 40{degrees}C, and no longer grew at 50{degrees}C. Legionella longbeachae reached a rather low cell density at 40{degrees}C and did not grow at and beyond 45{degrees}C. Genes encoding components of the Lqs-LvbR network were present in the genomes of the environmental and clinical L. pneumophila isolates, and the PlqsR, PlqsA, PlqsS and PlvbR promoters from strain JR32 were active in these strains. Taken together, our results indicate that the Lqs-LvbR network governs the temperature-dependent growth onset and cell density of the L. pneumophila reference strain JR32, and possibly also of environmental and clinical L. pneumophila isolates. ImportanceEnvironmental bacteria of the genus Legionella are the causative agents of the severe pneumonia Legionnaires disease, the incidence of which is worldwide on the rise. Legionella pneumophila and Legionella longbeachae are the clinically most relevant species. The opportunistic pathogens are inhaled through contaminated aerosols and replicate in human lung macrophages with a similar mechanism as in their natural hosts, free-living amoebae. Given their prevalence in natural and technical water systems, an efficient control of Legionella spp. by physical, chemical or biological means will reduce the incidence of Legionnaires disease. Here we report that the Legionella quorum sensing (Lqs) system and the pleiotropic transcription factor LvbR govern the temperature-dependent growth onset and cell density of bacterial cultures. Hence, the growth of L. pneumophila in water systems is not only determined by the temperature and nutrient availability, but also by quorum sensing, i.e., density- and signaling molecule-dependent gene regulation.