An immunoglobulin superfamily-like domain unique to the Yersinia pseudotuberculosis invasin protein is required for stimulation of bacterial uptake via integrin receptors.

The binding of the Yersinia pseudotuberculosis and Yersinia enterocolitica invasin proteins to beta(1) integrin receptors allows internalization of these organisms by cultured cells. The C-terminal 192-residue superdomain of the Y. pseudotuberculosis invasin is necessary and sufficient for integrin recognition, while a region located outside, and N-terminal to, this superdomain strongly enhances the efficiency of bacterial uptake. Within the enhancer region is a domain called D2 that allows invasin-invasin interaction. To investigate the role of the enhancer region, bacterial cell binding and entry mediated by the Y. pseudotuberculosis invasin protein (invasin(pstb)) was compared to that of Y. enterocolitica invasin (invasin(ent)), which lacks the D2 self-association domain. Invasin(ent) was shown to be unable to promote self-interaction, using the DNA binding domain of lambda repressor as a reporter. Furthermore, two genetically engineered in-frame deletion mutations that removed D2 from invasin(pstb) were significantly less proficient than wild-type invasin(pstb) at promoting uptake, although the amount of surface-exposed invasin as well as the cell binding capacity of the recombinant Escherichia coli strains remained similar. Competitive uptake assays showed that E. coli cells expressing invasin(pstb) had a significant advantage in the internalization process versus either E. coli cells expressing invasin(ent) or the invasin(pstb) derivatives deleted for D2, further demonstrating the importance of invasin self-interaction for the efficiency of invasin-mediated uptake.