Pseudomonas aeruginosa GacA, a factor in multihost virulence, is also essential for biofilm formation

We have investigated a potential role for GacA, the response regulator of t he GacA/GacS two-component regulatory system, in Pseudomonas aeruginosa bio film formation. When gacA was disrupted in strain PA14, a 10-fold reduction in biofilm formation capacity resulted relative to wild-type PA14. However , no significant difference was observed in the planktonic growth rate of P A14 gacA(-). Providing gacA in trans on the multicopy vector pUCP-gacA abro gated the biofilm formation defect. Scanning electron microscopy of biofilm s formed by PA14 gacA(-) revealed diffuse clusters of cells that failed to aggregate into microcolonies, implying a deficit in biofilm development or surface translocation. Motility assays revealed no decrease in PA14 gacA(-) twitching or swimming abilities, indicating that the defect in biofilm for mation is independent of flagellar-mediated attachment and solid surface tr anslocation by pili. Autoinducer and alginate bioassays were performed simi larly, and no difference in production levels was observed, indicating that this is not merely an upstream effect on either quorum sensing or alginate production. Antibiotic susceptibility profiling demonstrated that PA14 gac A(-) biofilms have moderately decreased resistance to a range of antibiotic s relative to PA14 wild type. This study establishes GacA as a new and inde pendent regulatory element in P. aeruginosa biofilm formation.