Abstract

The identification of cell-to-cell signaling inhibitors among bacteria is a novel mechanism used to control the virulence of pathogens. Signaling inhibitors have a significant potential to be used as therapeutics in that they will exert less biological pressure to develop resistance than classic antibiotics, which attempt to eradicate or significantly inhibit bacterial growth. Decreasing the virulence of pathogens by inhibiting signaling provides a mechanism by which the bacteria will be "misled" into not activating virulence factors. If virulence factors are not activated, these pathogens will either pass through the host without incidence or be controlled by the host's immune system. In this unit, we discuss the general principles for the design and implementation of a high-throughput screen to identify inhibitors of bacterial cell-to-cell communication. We also provide a detailed protocol using a specific example of signaling inhibitor identification using enterohemorrhagic Escherichia coli as a model system.