Pseudopaline-mediated zinc uptake by Pseudomonas aeruginosa determines specific clinically relevant phenotypes and infection outcome

AUTHOR SUMMARYThe host-pathogen interface is a biological niche in which two entities competes for essential resources. The hosts nutritional immunity restrict access to metals, while a successful pathogen overcomes these restrictions using dedicated uptake pathways. Pseudopaline is a high-affinity metallophore allowing Pseudomonas aeruginosa to acquire zinc in chelated environments. We demonstrate that this pathway is the last-resort solution to acquire zinc for this dreadful pathogen. The capacity to provide this metal to zinc-metalloproteins drives clinically relevant phenotypes, such as the capacity to form a mature and antibiotic-tolerant biofilm, or to affect the outcome of an infection. These results place pseudopaline as a potential drug target for blocking P. aeruginosa pathogenic capacity and resensitizing established biofilm to classic antibiotic treatment. ABSTRACTBiological metals are essential trace elements which are required by metalloproteins, involved in virtually every cellular, structural and catalytic function of the bacterial cell. Bacterial pathogenesis involves a tug-of-war between the host nutritional immunity, sequestering essential metals and the invading pathogens that deploy high-metal affinity uptake strategies in order to overcome these defence mechanisms. Metallophores are high-affinity, low-molecular mass metal chelators produced and secreted by bacteria to access chelated metals from the environment. Pseudopaline is a metallophore produced and secreted by Pseudomonas aeruginosa to acquire zinc when the bioavailability of this metal is severely restricted, as in the presence of a strong metal chelator such as EDTA, or during infections when the nutritional immunity of the host is active, in mammals through the production of the zing binding protein calprotectin. We show that under the conditions of metal deprivation, a pseudopaline-deficient P. aeruginosa strain exhibit a severe intracellular zinc deficiency, establishing that the pseudopaline pathway is the last-resort and unique pathway for the bacteria to acquire zinc under these restricted growth conditions. The present study explores the pleiotropic role of pseudopaline-mediated zinc acquisition on several clinically relevant phenotypes and its capacity to drive infection outcomes, placing this machinery as a promising therapeutic target for P. aeruginosas infection, acting synergistically as a pathogenicity determinant as well as an adaptative trait allowing the establishment of a mature and antibiotic resistance biofilm necessary for recalcitrant chronic infections.