Transcriptional regulation of prfA and PrfA‐regulated virulence genes in Listeria monocytogenes

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Abstract
The ActA protein, the lecithinase PlcB and listeriolysin are the major PrfA-dependant proteins synthesized when brain-heart infusion (BHI)-cultured Listeria monocytogenes is shifted to minimum essential medium (MEM) In the presence of the transcriptional inhibitor rifampicin. Enhanced synthesis of all three proteins under these conditions depends, however, on a short incubation (about 5 min) of the bacteria in MEM without rifampicin, suggesting that induction of these proteins in MEM requires de novo transcription. The enhanced synthesis of these three proteins is observed in the L. monocytogenes wild-type strains EGD and NCTC 7973, both of which belong to the serotype 1/2a. A significant induction of the bicistronic mRNA for ActA and PlcB is observed in both strains shortly after shifting the bacteria from BHI to MEM. This mRNA as well as the monocistronic listeriolysin (hly)-specific mRNA is highly stable in L. monocytogenes NCTC 7973 shifted to MEM. In contrast to the actA-plcB mRNA, no enhanced transcription in MEM is observed for the regulatory prfA gene or for the PrfA-controlled virulence genes hlyA and plcA in strain NCTC 7973. However, transcription of these genes is induced in strain EGD. Transcriptional induction of the mpl gene is observed in neither strain NCTC 7973 nor in strain EGD. The life-time of the prfA, plcA, and mpl transcripts is short. ActA was also found to be the most abundant newly synthesized surface protein when the two wild-type strains of L. monocytogenes replicated within the phagocytic cell line J774. ActA synthesis seemed to be induced in the cytoplasm since the non-haemolytic mutant M3 did not induce ActA when taken up by J774 cells.