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Journal of Bacteriology, May 2005, p. 3277-3281, Vol. 187, No. 9
0021-9193/05/$08.00+0     doi:10.1128/JB.187.9.3277-3281.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Novel Roles of ohrR-ohr in Xanthomonas Sensing, Metabolism, and Physiological Adaptive Response to Lipid Hydroperoxide

Chananat Klomsiri,1 Warunya Panmanee,2 Saovanee Dharmsthiti,3 Paiboon Vattanaviboon,2 and Skorn Mongkolsuk1*

Laboratory of Biotechnology, Chulabhorn Research Institute, Lak Si, Bangkok 10210, Thailand,2 Department of Biotechnology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand,1 Center for Biotechnology, Institute for Research and Development in Science and Technology, Mahidol University, Nakornprathom 73170, Thailand3

Received 17 September 2004/ Accepted 18 January 2005

Lipid hydroperoxides are highly toxic to biological systems. Here, the Xanthomonas campestris pv. phaseoli sensing and protective systems against linoleic hydroperoxide (LOOH) were investigated by examining the phenotypes, biochemical and regulatory characteristics of various Xanthomonas mutants in known peroxide resistance pathways. Analysis of LOOH resistance levels indicates that both alkyl hydroperoxide reductase (AhpC) and organic hydroperoxide resistance enzyme (Ohr) have important and nonredundant roles in the process. Nonetheless, inactivation of ohr leads to a marked reduction in LOOH resistance levels. The regulatory characteristics of an ohr mutant add further support to its primary role in LOOH protection. Northern analysis shows that LOOH had differential effects on induction of ahpC and ohr expression with the latter being more sensitive to the inducer. Analysis of the ahpC and ohr promoters confirmed that the LOOH-dependent induction of these promoters is mediated by the transcription regulators OxyR and OhrR, respectively. Using the in vivo promoter assays and the in vitro gel mobility shift assay, we show that LOOH directly oxidized OhrR at the sensing residue Cys-22 leading to its inactivation. In addition, physiological analysis shows that pretreatment of X. campestris pv. phaseoli with a sublethal dose of LOOH induced high levels of resistance to subsequent exposure to lethal concentrations of LOOH. This novel LOOH-induced adaptive response requires a functional ohrR-ohr operon. These data illustrate an important novel physiological role for the ohrR-ohr system in sensing and inactivating lipid hydroperoxides.

* Corresponding author. Mailing address: Laboratory of Biotechnology, Chulabhorn Research Institute, Lak Si, Bangkok 10210, Thailand. Phone: (662) 574 0630, ext. 3816. Fax: (662) 574 2027. E-mail: skorn{at}tubtim.cri.or.th.

Journal of Bacteriology, May 2005, p. 3277-3281, Vol. 187, No. 9
0021-9193/05/$08.00+0     doi:10.1128/JB.187.9.3277-3281.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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