A protease-resistant catalase, KatA, released upon cell lysis during stationary phase is essential for aerobic survival of a Pseudomonas aeruginosa oxyR mutant at low cell densities.
A Pseudomonas aeruginosa oxyR mutant was dramatically sensitive to H(2)O(2), despite possessing wild-type catalase activity. Oxygen-dependent oxyR phenotypes also included an inability to survive aerobic serial dilution in Luria broth and to resist aminoglycosides. Plating the oxyR mutant after serial dilution in its own spent culture supernatant, which contained the major catalase KatA, or under anaerobic conditions allowed for survival. KatA was resistant to sodium dodecyl sulfate, proteinase K, pepsin, trypsin, chymotrypsin and the neutrophil protease cathepsin G. When provided in trans and expressed constitutively, the OxyR-regulated genes katB, ahpB, and ahpCF could not restore both the serial dilution defect and H(2)O(2) resistance; only oxyR itself could do so. The aerobic dilution defect could be complemented, in part, by only ahpB and ahpCF, suggesting that the latter gene products could possess a catalase-like activity. Aerobic Luria broth was found to generate approximately 1.2 microM H(2)O(2) min(-1) via autoxidation, a level sufficient to kill serially diluted oxyR and oxyR katA bacteria and explain the molecular mechanism behind the aerobic serial dilution defect. Taken together, our results indicate that inactivation of OxyR renders P. aeruginosa exquisitely sensitive to both H(2)O(2) and aminoglycosides, which are clinically and environmentally important antimicrobials.
Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati College of Medicine, Cincinnati, Ohio 45267-0524, USA. Daniel.hassett@UC.Edu
D J
DJ
Hassett
E
E
Alsabbagh
K
K
Parvatiyar
M L
ML
Howell
R W
RW
Wilmott
U A
UA
Ochsner
D000332:Aerobiosis; D000693:Anaerobiosis; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002374:Catalase / Q000235:genetics / Q000378:metabolism*; D003470:Culture Media; D004268:DNA-Binding Proteins; D004352:Drug Resistance, Microbial; D005838:Genotype; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D016296:Mutagenesis; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D012097:Repressor Proteins / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics*
D000900:Anti-Bacterial Agents; D003470:Culture Media; D004268:DNA-Binding Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; D006861:Hydrogen Peroxide; D002374:Catalase