Architecture and function of cell wall synthesis complexes of Pseudomonas aeruginosa – PSEUDO-WALL

Pseudomonas aeruginosa is a causative agent of hospital- acquired infections, calling for the urgent development of new treatments. This grant builds on a very successful ANR project (ended Oct 2017) in which Partner 1 structurally and functionally characterized the first complex between a PBP and a scaffolding factor (MreC), key for bacterial cell wall elongation. Here, we will extend this work to characterize the cell wall assemblies of P. aeruginosa using crystallography, EM, biochemistry, and microbiology. In addition, we will search for inhibitors of the PBP:MreC interaction by fragment screening in a high throughput crystallization/data collection platform at the EMBL/ESRF in Grenoble, and identified ‘hits’ will be tested in microbial/transcriptomic assays by RNAseq. Lastly, we will employ a transposon library made in a clinical P. aeruginosa strain to identify additional partners and regulators of the P. aeruginosa elongasome, providing a fresh outlook onto the search for new antimicrobials.