Identification of Salmonella typhimurium invasiveness loci.
Salmonella typhimurium is capable of entering into (invading) nonphagocytic host cells. To systematically identify the bacterial genes necessary for this process, 15,000 Tn10dCm random transposon mutants of S. typhimurium were individually screened for invasiveness, using the human colonic epithelial Caco-2 cell line. Four hundred and eighty-eight mutants had decreased levels of invasiveness; most were nonmotile. However, five mutants, representing four loci, were completely motile. Further characterization of these five mutants showed that they were also unable to enter the dog kidney epithelial cell line MDCK and the mouse macrophage line J774.A1. In contrast to the parental strain, they were unable to disrupt the transepithelial resistance of polarized epithelial monolayers, nor were they able to penetrate across these epithelial barriers. Three of the four classes of mutants remained virulent in mice. The results confirm several aspects of S. typhimurium invasiveness: (i) intact motility enhances invasiveness of cultured cells; (ii) S. typhimurium invasiveness is multifactorial, and at least six distinct genetic loci are involved; and (iii) invasion loci involved in uptake into epithelial cells are also needed for uptake into cultured phagocytic cells. The results also emphasize that decreased levels of invasiveness eliminate bacterial penetration of polarized epithelial barriers and invasiveness loci mutants are not necessarily avirulent.
Betts
J
J
Department of Biochemistry, University of British Columbia, Vancouver, Canada.
Finlay
B B
BB
D000818:Animals; D002460:Cell Line; D002465:Cell Movement; D004252:DNA Mutational Analysis; D004269:DNA, Bacterial / Q000235:genetics*; D004285:Dogs; D051379:Mice; D012486:Salmonella typhimurium / Q000235:genetics* / Q000472:pathogenicity*; D014774:Virulence
D004269:DNA, Bacterial