title: A novel C-terminal mutation resulting in constitutive activation of the Listeria monocytogenes central virulence regulatory factor PrfA.
abstract: The environmental bacterium Listeria monocytogenes survives and replicates in a variety of diverse ecological niches that range from the soil to the cytosol of infected mammalian cells. The ability of L. monocytogenes to replicate within an infected host requires the expression of a number of secreted bacterial gene products whose expression is regulated by the transcriptional activator PrfA. PrfA becomes activated following bacterial entry into host cells; however, the mechanism by which this activation occurs remains unknown. Here we describe a novel C-terminal mutation that results in the high-level constitutive activation of PrfA and yet, in contrast with other described prfA* activation mutations, only modestly increases PrfA DNA binding affinity. L. monocytogenes strains containing the prfA P219S mutation exhibited high levels of PrfA-dependent virulence gene expression, were hyperinvasive in tissue culture models of infection, were fully motile and were hypervirulent in mice. In contrast with PrfA G145S and other mutationally activated PrfA proteins, the PrfA P219S protein readily formed homodimers and did not exhibit a dramatic increase in its DNA-binding affinity for target promoters. Interestingly, the prfA P219S mutation is located adjacent to the prfA K220 residue that has been previously reported to contribute to PrfA DNA binding activity. prfA P219S therefore appears to constitutively activate PrfA via a novel mechanism which minimally affects PrfA DNA binding in vitro.
authors.affiliation: Department of Microbiology and Immunology, University of Illinois at Chicago, Chicago, IL, USA.
authors.forename: Bobbi
authors.initials: B
authors.lastname: Xayarath
authors.affiliation: Department of Microbiology and Immunology, University of Illinois at Chicago, Chicago, IL, USA.
authors.forename: Jennifer I
authors.initials: JI
authors.lastname: Smart
authors.affiliation: Department of Microbiology and Immunology, University of Illinois at Chicago, Chicago, IL, USA.
authors.forename: Kimberly J
authors.initials: KJ
authors.lastname: Mueller
authors.affiliation: Department of Microbiology and Immunology, University of Illinois at Chicago, Chicago, IL, USA.
authors.forename: Nancy E
authors.initials: NE
authors.lastname: Freitag
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005260:Female; D015964:Gene Expression Regulation, Bacterial; D008089:Listeria monocytogenes / Q000235:genetics* / Q000472:pathogenicity; D051379:Mice; D009154:Mutation; D010454:Peptide Termination Factors / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D010454:Peptide Termination Factors; C545356:PrfA protein, Listeria monocytogenes; D037521:Virulence Factors