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Significance The virulence of numerous bacterial pathogens is dependent on nanomachines that inject tens of proteins (effectors) into host cells. Effectors are often functionally redundant or act in an additive manner to usurp cellular processes, limiting their characterization via traditional top-down loss-of-function assays. To overcome this issue, we developed an innovative synthetic bottom-up platform to conduct gain-of-function screens. These screens identified previously missedShigella effectors that inhibit induced epithelial cell death, an important arm of the host innate immune system triggered in response to invading pathogens. Our studies provide new mechanistic insight into the sophisticated strategies by whichShigella effectors interact with the host to enable this pathogen to establish a replicative niche within the cytosol of infected epithelial cells.

Synthetic bottom-up approach reveals the complex interplay of Shigella effectors in regulation of epithelial cell death