A suppressor of the menadione-hypersensitive phenotype of a Xanthomonas campestris pv. phaseoli oxyR mutant reveals a novel mechanism of toxicity and the protective role of alkyl hydroperoxide reductase.
We isolated menadione-resistant mutants of Xanthomonas campestris pv. phaseoli oxyR (oxyR(Xp)). The oxyRR2(Xp) mutant was hyperresistant to the superoxide generators menadione and plumbagin and was moderately resistant to H(2)O(2) and tert-butyl hydroperoxide. Analysis of enzymes involved in oxidative-stress protection in the oxyRR2(Xp) mutant revealed a >10-fold increase in AhpC and AhpF levels, while the levels of superoxide dismutase (SOD), catalase, and the organic hydroperoxide resistance protein (Ohr) were not significantly altered. Inactivation of ahpC in the oxyRR2(Xp) mutant resulted in increased sensitivity to menadione killing. Moreover, high levels of expression of cloned ahpC and ahpF in the oxyR(Xp) mutant complemented the menadione hypersensitivity phenotype. High levels of other oxidant-scavenging enzymes such as catalase and SOD did not protect the cells from menadione toxicity. These data strongly suggest that the toxicity of superoxide generators could be mediated via organic peroxide production and that alkyl hydroperoxide reductase has an important novel function in the protection against the toxicity of these compounds in X. campestris.
Vattanaviboon
Paiboon
P
Laboratory of Biotechnology, Chulabhorn Research Institute, Bangkok 10210, Thailand. paiboon@tubtim.cri.or.th
Whangsuk
Wirongrong
W
Mongkolsuk
Skorn
S
D001426:Bacterial Proteins / Q000187:drug effects / Q000378:metabolism; D002374:Catalase / Q000187:drug effects / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D008826:Microbial Sensitivity Tests; D009154:Mutation; D010544:Peroxidases / Q000187:drug effects / Q000502:physiology*; D054464:Peroxiredoxins; D013482:Superoxide Dismutase / Q000187:drug effects / Q000378:metabolism; D024483:Vitamin K 3 / Q000494:pharmacology* / Q000633:toxicity; D016959:Xanthomonas campestris / Q000187:drug effects* / Q000502:physiology*
D001426:Bacterial Proteins; C112257:organic hydroperoxide resistance protein, Xanthomonas campestris; D024483:Vitamin K 3; D010544:Peroxidases; D054464:Peroxiredoxins; D002374:Catalase; D013482:Superoxide Dismutase