Identification of a third EspA-binding protein that forms part of the type III secretion system of enterohemorrhagic Escherichia coli.

Enterohemorrhagic Escherichia coli utilizes a type III secretion system to deliver virulent effectors into cells. The secretion apparatus comprises a membrane basal body and an external needle complex of which EspA is the major component. An l0050-deletion (DeltaL50) mutation was found to impair type III secretion and bacterial adherence. These phenotypes and the localization of the gene product to the inner membrane support the hypothesis that L0050, renamed EscL, forms part of the secretion apparatus. Furthermore, in DeltaL50, the amount of EspA present within the cell lysate was found to have diminished, whereas the EspA co-cistron-expressed partner protein EspB remained unaffected. The decreased EspA level appeared to result from instability of the newly synthesized EspA protein in DeltaL50 rather than a decrease in EspA mRNA. Using both biochemical co-purification and a bacterial two-hybrid interaction system, we were able to conclude that EscL is a third protein that, in addition to CesAB and CesA2, interacts with EspA and enhances the stability of intracellular EspA.