title: Identification of EnvC and Its Cognate Amidases as Novel Determinants of Intrinsic Resistance to Cationic Antimicrobial Peptides.
abstract: Cationic antimicrobial peptides (CAMPs) are an essential part of the innate immune system. Some Gram-negative enteric pathogens, such asSalmonella enterica, show intrinsic resistance to CAMPs. However, the molecular basis of intrinsic resistance is poorly understood, largely due to a lack of information about the genes involved. In this study, using a microarray-based genomic technique, we screened the Keio collection of 3,985Escherichia colimutants for altered susceptibility to human neutrophil peptide 1 (HNP-1) and identifiedenvCandzapBas novel genetic determinants of intrinsic CAMP resistance. In CAMP killing assays, anE. coliΔenvCEcor ΔzapBEcmutant displayed a distinct profile of increased susceptibility to both LL-37 and HNP-1. Both mutants, however, displayed wild-type resistance to polymyxin B and human β-defensin 3 (HBD3), suggesting that the intrinsic resistance mediated by EnvC or ZapB is specific to certain CAMPs. A correspondingSalmonellaΔenvCSemutant showed similarly increased CAMP susceptibility. TheenvCmutants of bothE. coliandS. entericadisplayed increased surface negativity and hydrophobicity, which partly explained the increased CAMP susceptibility. However, the ΔenvCEcmutant, but not the ΔenvCSemutant, was defective in outer membrane permeability, excluding this defect as a common factor contributing to the increased CAMP susceptibility. Animal experiments showed that theSalmonellaΔenvCSemutant had attenuated virulence. Taken together, our results indicate that the role ofenvCin intrinsic CAMP resistance is likely conserved among Gram-negative enteric bacteria, demonstrate the importance of intrinsic CAMP resistance for full virulence ofS. enterica, and provide insight into distinct mechanisms of action of CAMPs.
authors.affiliation: Center for Biomolecular Sciences, College of Pharmacy, University of Illinois at Chicago, Chicago, Illinois, USA Department of Biopharmaceutical Sciences, College of Pharmacy, University of Illinois at Chicago, Chicago, Illinois, USA.
authors.forename: Tamiko
authors.initials: T
authors.lastname: Oguri
authors.affiliation: Department of Microbiology and Immunology, Indiana University School of Medicine-Northwest, Gary, Indiana, USA.
authors.forename: Won-Sik
authors.initials: WS
authors.lastname: Yeo
authors.affiliation: Department of Microbiology and Immunology, Indiana University School of Medicine-Northwest, Gary, Indiana, USA.
authors.forename: Taeok
authors.initials: T
authors.lastname: Bae
authors.affiliation: Center for Biomolecular Sciences, College of Pharmacy, University of Illinois at Chicago, Chicago, Illinois, USA Department of Biopharmaceutical Sciences, College of Pharmacy, University of Illinois at Chicago, Chicago, Illinois, USA hlee31@uic.edu.
authors.forename: Hyunwoo
authors.initials: H
authors.lastname: Lee
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000494:pharmacology; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D018797:Cell Cycle Proteins; D002463:Cell Membrane Permeability; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D010450:Endopeptidases / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity; D015870:Gene Expression; D057927:Hydrophobic and Hydrophilic Interactions; D008826:Microbial Sensitivity Tests; D009154:Mutation; D011112:Polymyxin B / Q000494:pharmacology; D019779:Salmonella enterica / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity; D055672:Static Electricity; D014774:Virulence; D023084:alpha-Defensins / Q000494:pharmacology*; D023083:beta-Defensins / Q000494:pharmacology; D054804:Cathelicidins
chemical_list: D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; D001426:Bacterial Proteins; D018797:Cell Cycle Proteins; C422950:DEFB103A protein, human; D023084:alpha-Defensins; D023083:beta-Defensins; C406413:human neutrophil peptide 1; D010450:Endopeptidases; D011112:Polymyxin B; D054804:Cathelicidins
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