Interaction of enterohemorrhagic Escherichia coli (EHEC) with mammalian cells: cell adhesion, type III secretion, and actin pedestal formation.
Infection by the food-borne pathogen enterohemorragic Escherichia coli (EHEC) can lead to diarrhea, hemorrhagic colitis, and, in the most serious cases, renal failure. A critical step in colonization is the unusual interaction between EHEC and the intestinal epithelium. EHEC is able to adhere to mammalian cells, and then, by injecting bacterial proteins, or "effectors," into the host cell via a type III secretion system, induces the formation of attaching and effacing (AE) lesions characterized by the accumulation of filamentous actin directly beneath bound bacteria. This unit describes methods to evaluate the ability of EHEC to adhere to cultured mammalian cells, to secrete type III effectors, and to promote the formation of actin "pedestals." These methods can not only be used to evaluate the contribution of specific EHEC gene products to adherence, type III secretion, and mammalian cell signaling, but also facilitate the analysis of the eukaryotic pathways controlling fundamental cellular processes such as actin assembly.
Savage
Pamela J
PJ
University of Massachusetts Medical School, Worcester, Massachusetts, USA.
Campellone
Kenneth G
KG
Leong
John M
JM
D000199:Actins / Q000378:metabolism*; D001422:Bacterial Adhesion; D002460:Cell Line; D054324:Enterohemorrhagic Escherichia coli / Q000472:pathogenicity* / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans
D000199:Actins; D029968:Escherichia coli Proteins