Loss of the Two-Component System TctD-TctE in Affects Biofilm Formation and Aminoglycoside Susceptibility in Response to Citric Acid.

The two-component system TctD-TctE is important for regulating the uptake of tricarboxylic acids in TctD-TctE accomplishes this through derepression of the gene , which encodes a tricarboxylic acid-specific porin. Previous work from our lab revealed that TctD-TctE in also has a role in resistance to aminoglycoside antibiotics. The aim of this study was to further characterize the role of TctD-TctE in in the presence of citric acid. Here it was found that deletion of PA14 TctD-TctE (Δ) resulted in a 4-fold decrease in the biofilm bactericidal concentrations of the aminoglycosides tobramycin and gentamicin when citric acid was present in nutrient media. Tobramycin accumulation assays demonstrated that deletion of TctD-TctE resulted in an increase in the amount of tobramycin retained in biofilm cells. The PA14 wild type responded to increasing concentrations of citric acid by producing less biofilm. In contrast, the amount of Δ mutant biofilm formation remained constant or enhanced. Furthermore, the Δ strain was incapable of growing on citric acid as a sole carbon source and was highly reduced in its ability to grow in the presence of citric acid even when an additional carbon source was available. Use of phenotypic and genetic microarrays found that this growth deficiency of the Δ mutant is unique to citric acid and that multiple metabolic genes are dysregulated. This work demonstrates that TctD-TctE in has a role in biofilm development that is dependent on citric acid and that is separate from the previously characterized involvement in resistance to antibiotics. Nutrient availability is an important contributor to the ability of bacteria to establish successful infections in a host. is an opportunistic pathogen in humans causing infections that are difficult to treat. In part, its success is attributable to a high degree of metabolic versatility. is able to sense and respond to varied and limited nutrient stress in the host environment. Two-component systems are important sensors-regulators of cellular responses to environmental stresses, such as those encountered in the host. This work demonstrates that the response by the two-component system TctD-TctE to the presence of citric acid has a role in biofilm formation, aminoglycoside susceptibility, and growth in .