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Milohanic, Eliane; Glaser, Philippe; Coppée, Jean‐Yves; Frangeul, Lionel; Vega, Yolanda; Vázquez‐Boland, José A.; Kunst, Frank; Cossart, Pascale; Buchrieser, Carmen
Molecular microbiology, March 2003, Letnik: 47, Številka: 6Journal Article
Summary PrfA is the major regulator of Listeria virulence gene expression. This protein is a member of the Crp/Fnr family of transcription regulators. To gain a deeper understanding of the PrfA regulon, we constructed a whole‐genome array based on the complete genome sequence of Listeria monocytogenes strain EGDe and evaluated the expression profiles of the wild‐type EGDe and a prfA‐deleted mutant (EGDe ΔprfA). Both strains were grown at 37°C in brain–heart infusion broth (BHI) and BHI supplemented with either activated charcoal, a compound known to enhance virulence gene expression, or cellobiose, a sugar reported to downregulate virulence gene expression in spite of full expression of PrfA. We identified three groups of genes that are regulated differently. Group I comprises, in addition to the 10 already known genes, two new genes, lmo2219 and lmo0788, both positively regulated and preceded by a putative PrfA box. Group II comprises eight negatively regulated genes: lmo0278 is preceded by a putative PrfA box, and the remaining seven genes (lmo0178–lmo0184) are organized in an operon. Group III comprises 53 genes, of which only two (lmo0596 and lmo2067) are preceded by a putative PrfA box. Charcoal addition induced upregulation of group I genes but abolished regulation by PrfA of most group III genes. In the presence of cellobiose, all the group I genes were downregulated, whereas group III genes remained fully activated. Group II genes were repressed in all conditions tested. A comparison of the expression profiles between a second L. monocytogenes strain (P14), its spontaneous mutant expressing a constitutively active PrfA variant (P14prfA*) and its corresponding prfA‐deleted mutant (P14ΔprfA) and the EGDe strain revealed interesting strain‐specific differences. Sequences strongly similar to a sigma B‐dependent promoter were identified upstream of 22 group III genes. These results suggest that PrfA positively regulates a core set of 12 genes preceded by a PrfA box and probably expressed from a sigma A‐dependent promoter. In contrast, a second set of PrfA‐regulated genes lack a PrfA box and are expressed from a sigma B‐dependent promoter. This study reveals that PrfA can act as an activator or a repressor and suggests that PrfA may directly or indirectly activate different sets of genes in association with different sigma factors.
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