Abstract 1410 Investigating Bacterial 2',3'-Cyclic Nucleotide Monophosphate Signaling

Novel intracellular signaling molecules, 2',3'-cyclic nucleotide monophosphates (2',3'-cNMPs), have been recently (re)discovered within both prokaryotes and eukaryotes. In plants and mammals, wounding has been found to cause increased levels of 2',3'-cNMPs. Our studies in bacteria suggest that 2',3'-cNMPs also are produced in response to cellular stress and that 2',3'-cNMP levels affect bacterial phenotypes, including biofilm formation, motility, and growth. The enzyme responsible for 2',3'-cNMP production in E. coli and Salmonella enterica has been identified and we have developed cellular tools to modulate 2',3'-cNMP levels in bacteria, allowing us to directly probe the effects of 2',3'-cNMP concentration. By modulating 2',3'-cNMP levels and quantifying changes in gene expression, we have identified additional pathways modulated by 2',3'-cNMPs and are investigating regulatory mechanisms controlling the transcriptional changes. In addition, we have generated 2',3'-cNMP-linked resins to isolate 2',3'-cNMP-binding proteins and identified ribosomal proteins, among others. In vitro studies demonstrated that 2',3'-cNMPs decrease translation, suggesting a direct mechanism for 2',3-cNMP-dependent control of bacterial phenotypes. Our current studies are focused on identifying 2',3'-cNMP binding sites, other proteins involved in 2',3'-cNMP metabolism and sensing, and the intersections of 2',3'-cNMP and other cyclic nucleotide signaling pathways. By dissecting the cellular roles of 2',3'-cNMPs within bacteria, these studies highlight novel pathways within prokaryotes and, in the future, potentially can be engineered to control bacterial behavior. The authors are grateful for funding of this work by the National Institutes of Health [R01GM125842 to E.E.W.] and Penn State University.