Z-form extracellular DNA is a structural component of the bacterial biofilm matrix

Biofilms are community architectures adopted by bacteria inclusive of a self-formed extracellular matrix that protects resident bacteria from diverse environmental stresses and, in many species, incorporates extracellular DNA (eDNA) and DNABII proteins for structural integrity throughout biofilm development. Here, we present evidence that this eDNA-based architecture relies on the rare Z-form. Z-form DNA accumulates as biofilms mature and, through stabilization by the DNABII proteins, confers structural integrity to the biofilm matrix. Indeed, substances known to drive B-DNA into Z-DNA promoted biofilm formation whereas those that drive Z-DNA into B-DNA disrupted extant biofilms. Importantly, we demonstrated that the universal bacterial DNABII family of proteins stabilizes both bacterial- and host-eDNA in the Z-form in situ. A model is proposed that incorporates the role of Z-DNA in biofilm pathogenesis, innate immune response, and immune evasion. [Display omitted] •Z-DNA is stable and abundant extracellularly•Z-DNA provides structure and DNase-resistance to the biofilm matrix•Bacterial DNABII proteins mitigate NET function The structural integrity of bacterial biofilms is strengthened by the action of bacterial DNABII proteins that drive the accumulation of extracellular, nuclease-resistant Z-form DNA and inhibit host neutrophil extracellular trap function.