Escherichia coli toxin gene hipA affects biofilm formation and DNA release.

Toxin-antitoxin (TA) systems in Escherichia coli may play a role in biofilm formation, but the mechanism involved remains debatable. It is not known whether the TA systems are responsible for extracellular DNA (eDNA) in biofilms. In this study, we investigated the function of the hipBA TA system in biofilm formation by Escherichia coli strain BW25113. First, the deletion of the HipBA TA system in E. coli BW25113 significantly reduced the biofilm biomass without antibiotic stress. Second, treatment of the BW25113 biofilm with DNase I caused a major reduction in biofilm formation, whereas similar treatment of the hipA mutant biofilm had only a minor effect. Third, the inactivation of HipA reduced the level of eDNA present in biofilm formation, and addition of BW25113 genomic DNA stimulated biofilm formation for both the wild-type and hipA mutant. Fourth, the wild-type cells underwent significantly more cell lysis than the hipA mutant. These results suggest that hipA plays a significant role during biofilm development and that eDNA is an important structural component of E. coli BW25113 biofilms. Thus, the TA system may enhance biofilm formation through DNA release.