title: Trafficking of porin-deficient Salmonella typhimurium mutants inside HeLa cells: ompR and envZ mutants are defective for the formation of Salmonella-induced filaments.
abstract: Outer membrane porin genes of Salmonella typhimurium, including ompC, ompF, and tppB, are regulated by the products of ompB, a two-component regulatory locus encoding OmpR and EnvZ. S. typhimurium ompR mutants are attenuated in mice, but to date no one has studied the intracellular trafficking of S. typhimurium porin-deficient mutants. In this study, isogenic transposon mutants of S. typhimurium with insertions in ompR, envZ, ompF, ompC, ompD, osmZ, and tppB were compared with wild-type SL1344 for trafficking in the human epithelial cell line HeLa. We found that ompR and envZ mutants were reduced or completely inhibited for the formation of Salmonella-induced filaments (Sifs). This result was confirmed with an ompB deletion mutant. Sifs are tubular structures containing lysosomal glycoprotein which are induced specifically by intracellular Salmonella. Genetic analysis showed that the ompR mutation could be complemented in trans by cloned ompR to restore its ability to induce Sifs. In contrast, mutations in the known ompR-regulated genes ompF, ompC, and tppB (as well as the ompR-independent porin gene, ompD) had no effect on Sif formation relative to that of wild-type SL1344, thus indicating that OmpR does not exert its role on these genes to induce Sif formation. The omp mutants studied were able to invade and replicate in HeLa cells at levels comparable to those in wild-type SL1344. We conclude that OmpR and EnvZ appear to regulate Sif formation triggered by intracellular S. typhimurium.
authors.affiliation: Biotechnology Laboratory, University of British Columbia, Vancouver, Canada.
authors.forename: S D
authors.initials: SD
authors.lastname: Mills
authors.forename: S R
authors.initials: SR
authors.lastname: Ruschkowski
authors.forename: M A
authors.initials: MA
authors.lastname: Stein
authors.forename: B B
authors.initials: BB
authors.lastname: Finlay
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D004251:DNA Transposable Elements; D029968:Escherichia coli Proteins; D006367:HeLa Cells / Q000382:microbiology; D006801:Humans; D009097:Multienzyme Complexes; D009154:Mutation; D018272:Porins / Q000032:analysis*; D012486:Salmonella typhimurium / Q000235:genetics / Q000502:physiology*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D004251:DNA Transposable Elements; D029968:Escherichia coli Proteins; D009097:Multienzyme Complexes; D018272:Porins; C057843:envZ protein, E coli