Characterization of transcription organization and analysis of unique expression patterns of an alkyl hydroperoxide reductase C gene (ahpC) and the peroxide regulator operon ahpF-oxyR-orfX from Xanthomonas campestris pv. phaseoli

Abstract

We have analyzed the transcription organization of ahpC, ahpF, oxyR, and orfX from Xanthomonas campestris pv. phaseoli. ahpC was transcribed as a monocistronic 0.6-kb mRNA, while ahpF-oxyR-orfX were transcribed as a pulycistronic approximately 3.0-kb-long mRNA. The novel transcription organization of these genes has not observed in other bacteria. Western analysis showed that oxidants (peroxides and superoxide anions), a thiol reagent (N-ethylmaleimide), and CdCl2caused large increases in the steady- state level of AhpC. Growth at alkaline pH also moderately induced AhpC accumulation. Thermal and osmotic stresses did not alter the levels of AhpC. Northern blotting results confirmed that oxidant- and CdCl2-induced AhpC accumulation was due to increased levels of ahpC transcripts. Analysis of oxyR expression revealed a unique pattern. Unlike other bacterial systems, peroxides and a superoxide generator induced accumulation of OxyR. Northern blotting results confirmed that these oxidants induced expression of oxyR operon. This novel regulatory pattern could be generally important. The transcription organization and patterns of chemicals and stress induction of ahpC and oxyR differed from those of other bacteria and are likely to be important for X. campestris pv. phaseoli survival during exposure to oxidants.