title: PrfA regulation offsets the cost of Listeria virulence outside the host.
abstract: Virulence traits are essential for pathogen fitness, but whether they affect microbial performance in the environment, where they are not needed, remains experimentally unconfirmed. We investigated this question with the facultative pathogen Listeria monocytogenes and its PrfA virulence regulon. PrfA-regulated genes are activated intracellularly (PrfA 'ON') but shut down outside the host (PrfA 'OFF'). Using a mutant PrfA regulator locked ON (PrfA*) and thus causing PrfA-controlled genes to be constitutively activated, we show that virulence gene expression significantly impairs the listerial growth rate (μ) and maximum growth (A) in rich medium. Deletion analysis of the PrfA regulon and complementation of a L. monocytogenes mutant lacking all PrfA-regulated genes with PrfA* indicated that the growth reduction was specifically due to the unneeded virulence determinants and not to pleiotropic regulatory effects of PrfA ON. No PrfA*-associated fitness disadvantage was observed in infected eukaryotic cells, where PrfA-regulated virulence gene expression is critical for survival. Microcosm experiments demonstrated that the constitutively virulent state strongly impaired L. monocytogenes performance in soil, the natural habitat of these bacteria. Our findings provide empirical proof that virulence carries a significant cost to the pathogen. They also experimentally substantiate the assumed, although not proven, key role of virulence gene regulation systems in suppressing the cost of bacterial virulence outside the host.
authors.affiliation: Microbial Pathogenesis Group, School of Biomedical Sciences, University of Edinburgh, Edinburgh, UK.|School of Biological Sciences, University of Edinburgh, Edinburgh, UK.
authors.forename: Radhakrishnan B
authors.initials: RB
authors.lastname: Vasanthakrishnan
authors.affiliation: Microbial Pathogenesis Group, School of Biomedical Sciences, University of Edinburgh, Edinburgh, UK.
authors.forename: Aitor
authors.initials: A
authors.lastname: de Las Heras
authors.affiliation: Microbial Pathogenesis Group, School of Biomedical Sciences, University of Edinburgh, Edinburgh, UK.|The Roslin Institute, University of Edinburgh, Edinburgh, UK.
authors.forename: Mariela
authors.initials: M
authors.lastname: Scortti
authors.affiliation: Microbial Pathogenesis Group, School of Biomedical Sciences, University of Edinburgh, Edinburgh, UK.
authors.forename: Caroline
authors.initials: C
authors.lastname: Deshayes
authors.affiliation: School of Biological Sciences, University of Edinburgh, Edinburgh, UK.|Centre for Immunity, Infection & Evolution, University of Edinburgh, Edinburgh, UK.
authors.forename: Nick
authors.initials: N
authors.lastname: Colegrave
authors.affiliation: Microbial Pathogenesis Group, School of Biomedical Sciences, University of Edinburgh, Edinburgh, UK.|The Roslin Institute, University of Edinburgh, Edinburgh, UK.|Centre for Immunity, Infection & Evolution, University of Edinburgh, Edinburgh, UK.|Grupo de Patogenómica Bacteriana, Universidad de León, León, Spain.
authors.forename: José A
authors.initials: JA
authors.lastname: Vázquez-Boland
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D006367:HeLa Cells; D054884:Host-Pathogen Interactions; D006801:Humans; D008089:Listeria monocytogenes / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity*; D010454:Peptide Termination Factors / Q000235:genetics*; D018085:Regulon; D012988:Soil Microbiology; D015534:Trans-Activators / Q000235:genetics; D037521:Virulence Factors / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D010454:Peptide Termination Factors; C545356:PrfA protein, Listeria monocytogenes; D015534:Trans-Activators; D037521:Virulence Factors