The role of charged molecules in the entry mechanism of enteroinvasive bacteria was studied using Escherichia coli HB101 harboring a plasmid (pRI203) containing the Yersinia pseudotuberculosis invasion region as an experimental model. We investigated the effect of several anionic and cationic polyelectrolytes on the initial steps of infection of HeLa S3 cells by E. coli HB101 (pRI203). Experiments in which the polyions were added to cell monolayers together with bacteria showed that invasion was only slightly influenced by anions whereas cations strongly enhanced bacterial entry. DEAE-dextran, histone and poly-L-lysine were the most effective enhancers producing an up to five-fold increase in the number of both infected cells and internalized bacteria. Moreover, addition of the active polycations at different stages of infection demonstrated that their action took place during the attachment step, whereas internalization was not affected.