Detailed examination of cytoskeletal networks within enteropathogenic Escherichia coli pedestals.

Enteropathogenic Escherichia coli (EPEC) manipulate the cytoskeleton of host intestinal epithelial cells, producing membrane protrusions termed pedestals that the bacteria reside on throughout the course of their infections. By definition pedestals are actin-based structures, however recent work has identified the spectrin cytoskeleton as a necessary component of EPEC pedestals. Here, we investigated the detailed arrangement of the spectrin and actin cytoskeletons within these structures. Immunofluorescent imaging revealed that the spectrin network forms a peripheral cage around actin at the membranous regions of pedestals. Myosin S1 fragment decorated actin filaments examined by electron microscopy demonstrated that actin filaments orientate with their fast-growing barbed ends toward the lateral membranes of EPEC pedestals. These findings provide a detailed descriptive analysis, which further illustrate the spectrin cytoskeletal organization within these structures.