structural and biochemical characterisation of the dual bpbp-seds peptidoglycan biosynthesis machinery in the intracellular pathogen klebsiella pneumo

"We have recently discovered that in a number of ESKAPE bacterial pathogens which adopt an intracellular mode of infection, including Klebsiella pneumoniae, a separate, intracellular specific PBP2 and PBP3 homology is encoded, although still associated with the cognate SEDs. Moreover, comparable work in Salmonella has revealed that the intracellular PBPs forms have entirely different b-lactam sensitivity which may underpin the ability of the pathogen to reinfect individuals infected with such pathogens, undermining clinical effectiveness of antibiotic treatment. Our hypothesis is that during K. pneumoniae intracellular infection, there must be a switch to the class b PBP required for intracellular infection, and that it is this protein which becomes part of an intracellular specific SEDS-PBP complex for division and elongation. A masters student in the Roper lab has established clones for all the possible combinations of RodA and FtsW with intracellular and extracellular forms of PBP2 and PBP3 respectively from K. pneumoniae which is one of the major ESKAPE pathogens of global concern. In this project we will investigate the structural and mechanistic biology of the respective SEDS-PBP combinations, leveraging our expertise in the biochemistry and structural biology of these proteins with specific reference to those in Klebsiella pneumoniae. This project will investigate this hypothesis and will be carried out as a collaboration with colleagues in Queens University Belfast (Professor Jose Bengoechea: https://pure.qub.ac.uk/en/persons/jose-bengoechea) who will perform complimentary in-vivo knock out experiments in bacterial cells and mammalian cell culture."