A central role for PBP2 in the activation of peptidoglycan polymerization by the bacterial cell elongation machinery

Cell elongation in rod-shaped bacteria is mediated by the Rod system, a conserved morphogenic complex that spatially controls cell wall assembly by the glycan polymerase RodA and crosslinking enzyme PBP2. Using Escherichia coli as a model system, we identified a PBP2 variant that promotes Rod system function when essential accessory components of the machinery are inactivated. This PBP2 variant hyperactivates cell wall synthesis in vivo and stimulates the activity of RodA-PBP2 complexes in vitro. Cells with the activated synthase also exhibited enhanced polymerization of the actin-like MreB component of the Rod system. Our results define an activation pathway governing Rod system function in which PBP2 conformation plays a central role in stimulating both glycan polymerization by its partner RodA and the formation of cytoskeletal filaments of MreB to orient cell wall assembly. In light of these results, previously isolated mutations that activate cytokinesis suggest that an analogous pathway may also control cell wall synthesis by the division machinery.Author summary: The cell wall of bacteria determines their shape and protects them from osmotic lysis. Two enzymatic activities are required for cell wall synthesis: glycan polymerization and crosslinking. A major new family of glycan polymerases was recently discovered and was proposed to work in complex with crosslinking enzymes called penicillin-binding proteins (PBPs). How the activities of these enzymes are coordinated to prevent the toxic generation of uncrosslinked glycans has remained unknown. Our analysis of the cell elongation system of Escherichia coli has revealed that this coupling is mediated by changes in the PBP that activate glycan chain synthesis by the polymerase. Furthermore, we present genetic evidence that this activation event is mediated by a component of the elongation machinery with a previously unknown function. Discovery of this activation pathway provides new mechanistic insight into the cell wall biogenesis process and identifies a new avenue to disrupt it for antibiotic development.