Differential expression of Listeria monocytogenes virulence genes in mammalian host cells.
We have used RT-PCR and GFP-mediated fluorescence to analyse the regulation of PrfA-dependent virulence genes of Listeria monocytogenes during proliferation in mammalian host cells. Our data show that most of the PrfA-regulated virulence genes are more efficiently expressed, as measured by transcript levels, when L. monocytogenes is grown in macrophages and macrophage-like cells rather than in epithelial cells, hepatocytes or endothelial cells. The promoters for hly and plcA are predominantly activated within the phagosomal compartment, while those for actA and inlC are predominantly activated in the host cell cytosol. Expression of actA and plcB precedes that of inlC after infection of epithelial cells and macrophages. Little transcription of inlA or inlB is observed in epithelial cells and there is only slightly more in macrophages. In both cell types the level of transcription of the inlAB operon is lower than is seen under extracellular growth conditions in rich media, which is compatible with the assumption that InlA and InlB are not required during intracellular growth of the bacteria. Activation of the PrfA-independent iap promoter is also low during intracellular growth, although the gene product (p60) is required for cell viability. The levels of the PrfA-dependent virulence gene transcripts do not correlate with the amount of prfA transcript present, which is low under all intracellular conditions analysed, suggesting that the prfA transcript is either highly unstable in bacteria that are growing intracellularly, or that the small amount of PrfA produced is highly activated by additional component(s).
Bubert
A
A
Lehrstuhl für Mikrobiologie, Theodor-Boveri-Institut für Biowissenschaften, Universität Würzburg, Germany. andreas.bubert@merck.de
Sokolovic
Z
Z
Chun
S K
SK
Papatheodorou
L
L
Simm
A
A
Goebel
W
W
D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D018938:Caco-2 Cells; D002451:Cell Compartmentation; D002460:Cell Line; D002478:Cells, Cultured; D005453:Fluorescence; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D049452:Green Fluorescent Proteins; D006801:Humans; D008089:Listeria monocytogenes / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity; D008164:Luminescent Proteins / Q000235:genetics; D008264:Macrophages / Q000166:cytology; D008322:Mammals; D051379:Mice; D008807:Mice, Inbred BALB C; D010454:Peptide Termination Factors; D010957:Plasmids; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015534:Trans-Activators / Q000378:metabolism*; D014158:Transcription, Genetic; D014774:Virulence
D001426:Bacterial Proteins; D008164:Luminescent Proteins; D010454:Peptide Termination Factors; D015534:Trans-Activators; D049452:Green Fluorescent Proteins