The Phosphoinositide Phosphatase SopB Manipulates Membrane Surface Charge and Trafficking of the Salmonella-Containing Vacuole

SummaryShifts in electrostatic surface charge of membranes have recently been highlighted as a significant factor contributing to protein targeting to the plasma membrane and nascent phagosomes. Intracellular, vacuole-adapted pathogens may also regulate surface charge of their vacuoles to establish a replicative niche. Since Salmonella enterica serovar Typhimurium controls trafficking of the Salmonella-containing vacuole (SCV) and inhibits its fusion with lysosomes, we investigated the contribution of surface charge to this process. Using recently developed fluorescent biosensors, we show that the bacterial phosphoinositide phosphatase SopB controls membrane surface charge of nascent SCVs by reducing levels of negatively charged lipids phosphatidylinositol-4,5-bisphosphate and phosphatidylserine. This SopB activity results in dissociation of a number of host-cell endocytic trafficking proteins from this compartment and inhibits SCV-lysosome fusion. Moreover, inducible reduction of negative charge rescues ΔsopB bacteria-containing SCVs from fusion with lysosomes. These results reveal a membrane-charge-based mechanism used by S. Typhimurium to control SCV maturation.

► S. Typhimurium effector SopB reduces negative membrane surface charge of nascent SCVs
► SopB reduces levels of phosphatidylserine in the SCV membrane
► SopB controls SCV targeting of Rabs that rely on electrostatics for localization
► SopB regulates SCV trafficking and inhibits SCV-lysosome fusion