We have previously observed that enteropathogenic Escherichia coli (EPEC) adhere to HeLa cells in a localized manner, which we designated localized adherence as opposed to the diffuse pattern of adhesion. In this paper we have examined the effects of localized adherence of EPEC on the actin microfilament system of host HeLa cells. Centrifugation of bacteria onto HeLa cells improved the localized adherence and rapid rearrangements of actin filaments were detected by immunofluorescence and electron microscopy. Aggregation of microfilaments is consistently observed at the sites of localized adherence, and is abolished by cytochalasin D and low temperatures. Scanning electron microscopy indicates that these aggregates are surface microvilli entangled with attached EPEC.