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

被引:84
作者
Hasset, DJ
Alsabbagh, E
Parvatiyar, K
Howell, ML
Wilmot, RW
Ochsner, UA
机构
[1] Univ Cincinnati, Coll Med, Dept Mol Genet Biochem & Microbiol, Cincinnati, OH 45267 USA
[2] Univ Cincinnati, Coll Med, Dept Pediat, Div Pulm Med Allergy & Clin Immunol, Cincinnati, OH 45267 USA
[3] Prot Express Inc, Cincinnati, OH 45219 USA
[4] Univ Colorado, Hlth Sci Ctr, Dept Microbiol, Denver, CO 80262 USA
关键词
D O I
10.1128/JB.182.16.4557-4563.2000
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
A Pseudomonas aeruginosa oxyR mutant was dramatically sensitive to H2O2, 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 H2O2 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 similar to 1.2 mu M H2O2 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 H2O2 and aminoglycosides, which are clinically and environmentally important antimicrobials.
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页码:4557 / 4563
页数:7
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