title: Two-pronged survival strategy for the major cystic fibrosis pathogen, Pseudomonas aeruginosa, lacking the capacity to degrade nitric oxide during anaerobic respiration.
abstract: Protection from NO gas, a toxic byproduct of anaerobic respiration in Pseudomonas aeruginosa, is mediated by nitric oxide (NO) reductase (NOR), the norCB gene product. Nevertheless, a norCB mutant that accumulated approximately 13.6 microM NO paradoxically survived anaerobic growth. Transcription of genes encoding nitrate and nitrite reductases, the enzymes responsible for NO production, was reduced >50- and 2.5-fold in the norCB mutant. This was due, in part, to a predicted compromise of the [4Fe-4S](2+) cluster in the anaerobic regulator ANR by physiological NO levels, resulting in an inability to bind to its cognate promoter DNA sequences. Remarkably, two O(2)-dependent dioxygenases, homogentisate-1,2-dioxygenase (HmgA) and 4-hydroxyphenylpyruvate dioxygenase (Hpd), were derepressed in the norCB mutant. Electron paramagnetic resonance studies showed that HmgA and Hpd bound NO avidly, and helped protect the norCB mutant in anaerobic biofilms. These data suggest that protection of a P. aeruginosa norCB mutant against anaerobic NO toxicity occurs by both control of NO supply and reassignment of metabolic enzymes to the task of NO sequestration.
authors.affiliation: Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati College of Medicine, Cincinnati, OH 45267-0524, USA.
authors.forename: Sang Sun
authors.initials: SS
authors.lastname: Yoon
authors.forename: Ahmet C
authors.initials: AC
authors.lastname: Karabulut
authors.forename: John D
authors.initials: JD
authors.lastname: Lipscomb
authors.forename: Robert F
authors.initials: RF
authors.lastname: Hennigan
authors.forename: Sergei V
authors.initials: SV
authors.lastname: Lymar
authors.forename: Stephanie L
authors.initials: SL
authors.lastname: Groce
authors.forename: Andrew B
authors.initials: AB
authors.lastname: Herr
authors.forename: Michael L
authors.initials: ML
authors.lastname: Howell
authors.forename: Patricia J
authors.initials: PJ
authors.lastname: Kiley
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Schurr
authors.forename: Benjamin
authors.initials: B
authors.lastname: Gaston
authors.forename: Kyoung-Hee
authors.initials: KH
authors.lastname: Choi
authors.forename: Herbert P
authors.initials: HP
authors.lastname: Schweizer
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Hassett
mesh_terms: D000693:Anaerobiosis; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology; D003550:Cystic Fibrosis / Q000382:microbiology*; D015180:Electrophoresis, Gel, Two-Dimensional; D009154:Mutation; D009569:Nitric Oxide / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000254:growth & development / Q000378:metabolism / Q000502:physiology*; D013057:Spectrum Analysis / Q000379:methods; D014158:Transcription, Genetic
chemical_list: D001426:Bacterial Proteins; D009569:Nitric Oxide