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Investigation of NO‐mediated synthesis of c‐di‐GMP and its role in bacterial biofilm formation

Bacteria colonize most surfaces, forming multicellular communities called biofilms. Biofilms are estimated to be responsible for up to 80% of human disease. The signaling molecule nitric oxide (NO) has been implicated in biofilms through an unknown pathway. In the genomes of many bacteria, an H‐NOX (Heme Nitric oxide/OXygen binding domain) is found next to a di‐guanylate cyclase (DGC) protein. H‐NOX domains define a new family of putative NO sensing proteins. The product of DGC proteins, cyclic di‐GMP, is a bacterial second messenger that is implicated in regulating the motility/biofilm growth switch in bacteria. We propose that H‐NOX responds to NO through the DGC, resulting in biofilm formation. We have characterized the DGC and determined the effect of NO/H‐NOX on cyclase activity. Additionally, we are detailing the effects of NO/H‐NOX on biofilm development via biofilm growth studies and genetic manipulations.