title: Structural basis for glutathione-mediated activation of the virulence regulatory protein PrfA in Listeria.
abstract: Infection by the human bacterial pathogen Listeria monocytogenes is mainly controlled by the positive regulatory factor A (PrfA), a member of the Crp/Fnr family of transcriptional activators. Published data suggest that PrfA requires the binding of a cofactor for full activity, and it was recently proposed that glutathione (GSH) could fulfill this function. Here we report the crystal structures of PrfA in complex with GSH and in complex with GSH and its cognate DNA, the hly operator PrfA box motif. These structures reveal the structural basis for a GSH-mediated allosteric mode of activation of PrfA in the cytosol of the host cell. The crystal structure of PrfAWTin complex only with DNA confirms that PrfAWTcan adopt a DNA binding-compatible structure without binding the GSH activator molecule. By binding to PrfA in the cytosol of the host cell, GSH induces the correct fold of the HTH motifs, thus priming the PrfA protein for DNA interaction.
authors.affiliation: Department of Chemistry, Umeå University, 901 87 Umeå, Sweden.|Umeå Centre for Microbial Research, Umeå University, 901 87 Umeå, Sweden.
authors.forename: Michael
authors.initials: M
authors.lastname: Hall
authors.affiliation: Department of Chemistry, Umeå University, 901 87 Umeå, Sweden.|Umeå Centre for Microbial Research, Umeå University, 901 87 Umeå, Sweden.
authors.forename: Christin
authors.initials: C
authors.lastname: Grundström
authors.affiliation: Department of Chemistry, Umeå University, 901 87 Umeå, Sweden.|Umeå Centre for Microbial Research, Umeå University, 901 87 Umeå, Sweden.
authors.forename: Afshan
authors.initials: A
authors.lastname: Begum
authors.affiliation: Department of Chemistry, Umeå University, 901 87 Umeå, Sweden.|Umeå Centre for Microbial Research, Umeå University, 901 87 Umeå, Sweden.
authors.forename: Mikael J
authors.initials: MJ
authors.lastname: Lindberg
authors.affiliation: Department of Chemistry, Umeå University, 901 87 Umeå, Sweden.|Umeå Centre for Microbial Research, Umeå University, 901 87 Umeå, Sweden.
authors.forename: Uwe H
authors.initials: UH
authors.lastname: Sauer
authors.affiliation: Department of Chemistry, Umeå University, 901 87 Umeå, Sweden.|Umeå Centre for Microbial Research, Umeå University, 901 87 Umeå, Sweden.
authors.forename: Fredrik
authors.initials: F
authors.lastname: Almqvist
authors.affiliation: Umeå Centre for Microbial Research, Umeå University, 901 87 Umeå, Sweden.|Department of Molecular Biology, Umeå University, 901 87 Umeå, Sweden.|Molecular Infection Medicine Sweden, Umeå University, 901 87 Umeå, Sweden.
authors.forename: Jörgen
authors.initials: J
authors.lastname: Johansson
authors.affiliation: Department of Chemistry, Umeå University, 901 87 Umeå, Sweden; elisabeth.sauer-eriksson@umu.se.|Umeå Centre for Microbial Research, Umeå University, 901 87 Umeå, Sweden.
authors.forename: A Elisabeth
authors.initials: AE
authors.lastname: Sauer-Eriksson
mesh_terms: D020816:Amino Acid Motifs; D001426:Bacterial Proteins / Q000378:metabolism*; D018360:Crystallography, X-Ray; D004269:DNA, Bacterial / Q000737:chemistry; D015964:Gene Expression Regulation, Bacterial; D005978:Glutathione / Q000737:chemistry; D005998:Glycine / Q000737:chemistry; D008089:Listeria monocytogenes / Q000378:metabolism*; D010454:Peptide Termination Factors / Q000378:metabolism*; D011485:Protein Binding; D055503:Protein Multimerization; D015534:Trans-Activators / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D010454:Peptide Termination Factors; C545356:PrfA protein, Listeria monocytogenes; D015534:Trans-Activators; D014157:Transcription Factors; D005978:Glutathione; D005998:Glycine
keywords: Listeria; PrfA; activation; glutathione; virulence