SigmaB contributes to Listeria monocytogenes invasion by controlling expression of inlA and inlB.
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 Delta sigB 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 sigmaB, 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 sigmaB contributes to both InlA and InlB-mediated invasion of L. monocytogenes. Previous identification of the sigmaB-dependent P2(prfA) promoter upstream of the major virulence gene regulator, positive regulatory factor A (PrfA), suggested that the contributions of sigmaB to expression of various virulence genes, including inlA, could be at least partially mediated through PrfA. To test this hypothesis, relative invasion capabilities of Delta sigB and Delta prfA strains were compared. Exponential-phase cells of the Delta sigB and Delta prfA strains were similarly defective at invasion; however, stationary-phase Delta sigB cells were significantly less invasive than stationary-phase DeltaprfA cells, suggesting that the contributions of sigmaB 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 sigmaB-dependent manner in stationary-phase L. monocytogenes. Together, results from this study provide strong biological evidence of a critical role for sigmaB in L. monocytogenes invasion into non-phagocytic cells, primarily mediated through control of inlA and inlB expression.
Kim
Heesun
H
Department of Food Science, Cornell University, Ithaca, NY 14853, USA.
Marquis
Hélène
H
Department of Microbiology and Immunology, Cornell University, Ithaca, NY 14853, USA.
Boor
Kathryn J
KJ
Department of Food Science, Cornell University, Ithaca, NY 14853, USA.
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D018938:Caco-2 Cells; D002460:Cell Line; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D008089:Listeria monocytogenes / Q000235:genetics / Q000254:growth & development / Q000378:metabolism / Q000472:pathogenicity*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D012808:Sigma Factor / Q000378:metabolism*
D001426:Bacterial Proteins; D008565:Membrane Proteins; C079011:SigB protein, Bacteria; D012808:Sigma Factor; C095768:inlB protein, Listeria monocytogenes; C069761:internalin protein, Bacteria