The mechanistic basis of the membrane‐permeabilizing activities of the virulence‐associated protein A (VapA) from Rhodococcus equi

Abstract

Pathogenic Rhodococcus equi release the virulence‐associated protein A (VapA) within macrophage phagosomes. VapA permeabilizes phagosome and lysosome membranes and reduces acidification of both compartments. Using biophysical techniques, we found that VapA interacts with model membranes in four steps: (i) binding, change of mechanical properties, (ii) formation of specific membrane domains, (iii) permeabilization within the domains, and (iv) pH‐specific transformation of domains. Biosensor data revealed that VapA binds to membranes in one step at pH 6.5 and in two steps at pH 4.5 and decreases membrane fluidity. The integration of VapA into lipid monolayers was only significant at lateral pressures <20 mN m−1 indicating preferential incorporation into membrane regions with reduced integrity. Atomic force microscopy of lipid mono‐ and bilayers showed that VapA increased the surface heterogeneity of liquid disordered domains. Furthermore, VapA led to the formation of a new microstructured domain type and, at pH 4.5, to the formation of 5 nm high domains. VapA binding, its integration and lipid domain formation depended on lipid composition, pH, protein concentration and lateral membrane pressure. VapA‐mediated permeabilization is clearly distinct from that caused by classical microbial pore formers and is a key contribution to the multiplication of Rhodococcus equi in phagosomes.

Mechanisms of host cell exit by intracellular pathogens

“…In addition, two reviews summarize apicomplexan exit strategies (Jimenéz-Ruiz et al, 2023;Scheiner et al, 2023). Three more research articles and a review deal with the lytic vacuolar exit by the bacterial pathogens Legionella (Hiller et al, 2023;Neuber et al, 2023), Rhodococcus (Nehls et al, 2024), Salmonella (Scharte et al, 2023), and Staphylococcus (Bayer et al, 2023).…”