The ability of Listeria monocytogenes to invade non-phagocytic cells is important for development of a systemic listeriosis infection. The authors previously reported that a L. monocytogenes ΔsigB strain is defective in invasion into human intestinal epithelial cells, in part, due to decreased expression of a major invasion gene, inlA. To characterize additional invasion mechanisms under the control of σB, mutants were generated carrying combinations of in-frame deletions in inlA, inlB and sigB. Quantitative assessment of bacterial invasion into the human enterocyte Caco-2 and hepatocyte HepG-2 cell lines demonstrated that σB contributes to both InlA and InlB-mediated invasion of L. monocytogenes. Previous identification of the σB-dependent P2prfA promoter upstream of the major virulence gene regulator, positive regulatory factor A (PrfA), suggested that the contributions of σB to expression of various virulence genes, including inlA, could be at least partially mediated through PrfA. To test this hypothesis, relative invasion capabilities of ΔsigB and ΔprfA strains were compared. Exponential-phase cells of the ΔsigB and ΔprfA strains were similarly defective at invasion; however, stationary-phase ΔsigB cells were significantly less invasive than stationary-phase ΔprfA cells, suggesting that the contributions of σB to invasion extend beyond those mediated through PrfA in stationary-phase L. monocytogenes. TaqMan quantitative reverse-transcriptase PCRs further demonstrated that expression of inlA and inlB was greatly increased in a σB-dependent manner in stationary-phase L. monocytogenes. Together, results from this study provide strong biological evidence of a critical role for σB in L. monocytogenes invasion into non-phagocytic cells, primarily mediated through control of inlA and inlB expression.
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