Host CD59 potentiates the type III secretion system in Yersinia pseudotuberculosis

Abstract

Type III secretion systems (T3SS) play critical roles in disease for many Gram-negative pathogens. Although the structural details of T3SS assembly have been extensively studied, the intimate interactions between T3SS components and the target host plasma membrane are poorly characterized. A previous RNA interference (RNAi) study identified the GPI-linked surface protein CD59 as a critical host component that supports the translocation of Yersinia pseudotuberculosis ( Yptb ) T3SS effectors into the host cytosol. We show here that both depletion and increased levels of CD59 reduced Yptb T3SS pore formation and Yop translocation into host target cells. CD59 is unlikely to act as a receptor for the T3SS translocon, as physical blockade of CD59 or enzymatic removal of all GPI-linked surface proteins had no effect on T3SS function. Consistent with its importance in supporting T3SS function, efficient bacterial effector-mediated focal adhesion disassembly was dependent on CD59. Surprisingly, depletion of CD59 interfered with the rearrangement of the lipid raft marker GM1 that occurs after Yptb adhesion to host cells, indicating that CD59 supports lipid microdomain dynamics. Consistent with a role in modulating lipid composition, the loss of CD59 led to the accumulation of phosphatidylcholine and alterations in both fatty acid chain lengths and saturation levels. This work points to a role for CD59 in maintaining plasma membrane dynamics necessary for efficient T3SS translocon function. In addition, a polarization index is described that quantifies the asymmetric association of membrane compartments with organelles.