Light emission from a Mudlux transcriptional fusion in Salmonella typhimurium is stimulated by hydrogen peroxide and by interaction with the mouse macrophage cell line J774.2.
Hydrogen peroxide is known to induce a multigenic response in Salmonella typhimurium cells. We have used a Mudlux transcriptional reporter system to identify and isolate fusions in the virulent strain SL1344 which respond to hydrogen peroxide in vitro by light production, and one of these fusions, MPG203, has been further characterized. Transient light production was observed from MPG203 at levels of hydrogen peroxide as low as 10 microM. However, high levels of this toxic oxidizing agent resulted in light suppression, particularly at low bacterial densities. This fusion was also shown to produce light following adhesion to cells of the mouse macrophage cell line J774.2. Furthermore, the response was greatly reduced in the presence of catalase, directly implicating hydrogen peroxide as the eliciting agent and suggesting the involvement of the hydrogen peroxide-induced bacterial stress response in the infection process. Chemiluminescence studies also indicated that inhibition of the respiratory burst may occur as the infection ratio is increased. In addition, the level of light produced from bacteria within individual macrophage cells was shown to vary.
Francis
K P
KP
Institute of Cell and Molecular Biology, University of Edinburgh, Scotland, United Kingdom.
Gallagher
M P
MP
D000818:Animals; D010583:Bacteriophage mu / Q000235:genetics*; D002374:Catalase / Q000494:pharmacology; D002460:Cell Line; D003001:Cloning, Molecular; D005839:Gentamicins / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D008027:Light; D008264:Macrophages / Q000382:microbiology*; D051379:Mice; D016897:Respiratory Burst; D012486:Salmonella typhimurium / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D013696:Temperature; D014158:Transcription, Genetic
D005839:Gentamicins; D006861:Hydrogen Peroxide; D002374:Catalase