TraDIS_investigation_of_the_full_effector_and_immunity_protein_repertoire_and_target_cell_receptor_components_for_the_type_VI_secretion_system_of_p__aeruginosa

In a previously very successful collaboration with the WTSI we have used TraDIS to identify known and novel H1-T6SS associated toxin-immunity pairs. Additionally, this work was able to identify genes which may have a protective role for the cell against T6SS targeting. While the receptor target for other similar contact-dependent systems has been identified and characterised, this has been neglected in the T6SS field. We hypothesis that the genes identified through TraDIS which provide protection against H1-T6SS mediated killing, act as T6SS receptors on the target cell. We are currently in the process of fully characterising the novel putative toxin-immunity pairs and putative receptor targets which we have identified in this work. In the current proposal we will extend this work to identify novel H2- and H3-T6SS associated toxin-immunity genes and receptor targets in P. aerguinosa. Additionally, we will generate TraDIS libraries in E. coli and P. putida and compete these libraries against T6SS active or inactive P. aeruginosa attacking strains. The aim of this work will be to recover E. coli and P. putida mutants which lack the T6SS target receptor(s), as these should not be killed in the competition assay against the active T6SS strain.