title: Attenuating Listeria monocytogenes Virulence by Targeting the Regulatory Protein PrfA.
abstract: The transcriptional activator PrfA, a member of the Crp/Fnr family, controls the expression of some key virulence factors necessary for infection by the human bacterial pathogen Listeria monocytogenes. Phenotypic screening identified ring-fused 2-pyridone molecules that at low micromolar concentrations attenuate L. monocytogenes cellular uptake by reducing the expression of virulence genes. These inhibitors bind the transcriptional regulator PrfA and decrease its affinity for the consensus DNA-binding site. Structural characterization of this interaction revealed that one of the ring-fused 2-pyridones, compound 1, binds at two separate sites on the protein: one within a hydrophobic pocket or tunnel, located between the C- and N-terminal domains of PrfA, and the second in the vicinity of the DNA-binding helix-turn-helix motif. At both sites the compound interacts with residues important for PrfA activation and helix-turn-helix formation. Ring-fused 2-pyridones represent a new class of chemical probes for studying virulence in L. monocytogenes.
authors.affiliation: Department of Chemistry, Umeå University, 901 87 Umeå, Sweden; Umeå Centre for Microbial Research (UCMR), Umeå University, 901 87 Umeå, Sweden.
authors.forename: James A D
authors.initials: JA
authors.lastname: Good
authors.affiliation: Umeå Centre for Microbial Research (UCMR), Umeå University, 901 87 Umeå, Sweden; Department of Molecular Biology, Umeå University, 901 87 Umeå, Sweden; Molecular Infection Medicine, Sweden (MIMS), Umeå University, 901 87 Umeå, Sweden.
authors.forename: Christopher
authors.initials: C
authors.lastname: Andersson
authors.affiliation: Umeå Centre for Microbial Research (UCMR), Umeå University, 901 87 Umeå, Sweden; Department of Molecular Biology, Umeå University, 901 87 Umeå, Sweden; Molecular Infection Medicine, Sweden (MIMS), Umeå University, 901 87 Umeå, Sweden.
authors.forename: Sabine
authors.initials: S
authors.lastname: Hansen
authors.affiliation: Umeå Centre for Microbial Research (UCMR), Umeå University, 901 87 Umeå, Sweden; Department of Molecular Biology, Umeå University, 901 87 Umeå, Sweden; Molecular Infection Medicine, Sweden (MIMS), Umeå University, 901 87 Umeå, Sweden.
authors.forename: Jessica
authors.initials: J
authors.lastname: Wall
authors.affiliation: Department of Chemistry, Umeå University, 901 87 Umeå, Sweden; Umeå Centre for Microbial Research (UCMR), Umeå University, 901 87 Umeå, Sweden.
authors.forename: K Syam
authors.initials: KS
authors.lastname: Krishnan
authors.affiliation: Department of Chemistry, Umeå University, 901 87 Umeå, Sweden; Umeå Centre for Microbial Research (UCMR), 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 (UCMR), 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.
authors.forename: Moritz S
authors.initials: MS
authors.lastname: Niemiec
authors.affiliation: Umeå Centre for Microbial Research (UCMR), Umeå University, 901 87 Umeå, Sweden; Department of Molecular Biology, Umeå University, 901 87 Umeå, Sweden; Molecular Infection Medicine, Sweden (MIMS), Umeå University, 901 87 Umeå, Sweden.
authors.forename: Karolis
authors.initials: K
authors.lastname: Vaitkevicius
authors.affiliation: Department of Chemistry, Umeå University, 901 87 Umeå, Sweden; Umeå Centre for Microbial Research (UCMR), Umeå University, 901 87 Umeå, Sweden.
authors.forename: Erik
authors.initials: E
authors.lastname: Chorell
authors.affiliation: Department of Chemistry, Umeå University, 901 87 Umeå, Sweden.
authors.forename: Pernilla
authors.initials: P
authors.lastname: Wittung-Stafshede
authors.affiliation: Department of Chemistry, Umeå University, 901 87 Umeå, Sweden; Umeå Centre for Microbial Research (UCMR), 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 (UCMR), Umeå University, 901 87 Umeå, Sweden. Electronic address: elisabeth.sauer-eriksson@umu.se.
authors.forename: A Elisabeth
authors.initials: AE
authors.lastname: Sauer-Eriksson
authors.affiliation: Department of Chemistry, Umeå University, 901 87 Umeå, Sweden; Umeå Centre for Microbial Research (UCMR), Umeå University, 901 87 Umeå, Sweden. Electronic address: fredrik.almqvist@umu.se.
authors.forename: Fredrik
authors.initials: F
authors.lastname: Almqvist
authors.affiliation: Umeå Centre for Microbial Research (UCMR), Umeå University, 901 87 Umeå, Sweden; Department of Molecular Biology, Umeå University, 901 87 Umeå, Sweden; Molecular Infection Medicine, Sweden (MIMS), Umeå University, 901 87 Umeå, Sweden. Electronic address: jorgen.johansson@umu.se.
authors.forename: Jörgen
authors.initials: J
authors.lastname: Johansson
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites / Q000187:drug effects; D018938:Caco-2 Cells; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D006367:HeLa Cells; D006801:Humans; D008089:Listeria monocytogenes / Q000187:drug effects* / Q000472:pathogenicity*; D008958:Models, Molecular; D010454:Peptide Termination Factors / Q000235:genetics / Q000378:metabolism*; D011728:Pyridones / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D014774:Virulence / Q000187:drug effects
chemical_list: D001426:Bacterial Proteins; D010454:Peptide Termination Factors; C545356:PrfA protein, Listeria monocytogenes; D011728:Pyridones; C017679:2-hydroxypyridine