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Infect Immun, April 1998, p. 1806-1811, Vol. 66, No. 4
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Trafficking of Porin-Deficient Salmonella typhimurium Mutants inside HeLa Cells: ompR and envZ Mutants Are Defective for the Formation of Salmonella-Induced Filaments

Scott D. Mills,dagger Sharon R. Ruschkowski, Murry A. Stein,Dagger and B. Brett Finlay*

Biotechnology Laboratory, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada

Received 17 July 1997/Returned for modification 1 September 1997/Accepted 14 January 1998

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.

* Corresponding author. Mailing address: Biotechnology Laboratory, Room 237, Wesbrook Building, 6174 University Blvd., Vancouver, British Columbia V6T 1Z3, Canada. Phone: (604) 822-2493. Fax: (604) 822-9830. E-mail: bfinlay{at}unixg.ubc.ca.

dagger Present address: Astra Research Center Boston, Inc., Cambridge, MA 02139-4239.

Dagger Present address: Department of Microbiology and Molecular Genetics, University of Vermont, Burlington, VT 05405-0068.




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