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Journal of Bacteriology, August 2001, p. 4405-4412, Vol. 183, No. 15
0021-9193/01/$04.00+0 DOI: 10.1128/JB.183.15.4405-4412.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Complex Regulation of the Organic Hydroperoxide
Resistance Gene (ohr) from Xanthomonas
Involves OhrR, a Novel Organic Peroxide-Inducible Negative
Regulator, and Posttranscriptional Modifications
Rojana
Sukchawalit,1,
Suvit
Loprasert,1
Sopapan
Atichartpongkul,1 and
Skorn
Mongkolsuk1,2,*
Laboratory of Biotechnology, Chulabhorn
Research Institute, Lak Si, Bangkok 10210,1 and
Department of Biotechnology, Faculty of Science, Mahidol
University, Bangkok 10400,2 Thailand
Received 14 February 2001/Accepted 1 May 2001
Analysis of the sequence immediate upstream of ohr
revealed an open reading frame, designated ohrR, with
the potential to encode a 17-kDa peptide with moderate amino acid
sequence homology to the MarR family of negative regulators of gene
expression. ohrR was transcribed as bicistronic mRNA
with ohr, while ohr mRNA was found to be
95% monocistronic and 5% bicistronic with ohrR. Expression of both genes was induced by tert-butyl
hydroperoxide (tBOOH) treatment. High-level expression of
ohrR negatively regulated ohr expression.
This repression could be overcome by tBOOH treatment. In vivo promoter
analysis showed that the ohrR promoter (P1) has organic
peroxide-inducible, strong activity, while the ohr
promoter (P2) has constitutive, weak activity. Only P1 is autoregulated by OhrR. ohr primer extension results revealed three
major primer extension products corresponding to the 5' ends of
ohr mRNA, and their levels were strongly induced by
tBOOH treatment. Sequence analysis of regions upstream of these sites
showed no typical Xanthomonas promoter. Instead, the
regions can form a stem-loop secondary structure with the 5' ends of
ohr mRNA located in the loop section. The secondary
structure resembles the structure recognized and processed by RNase III
enzyme. These findings suggest that the P1 promoter is responsible for
tBOOH-induced expression of the ohrR-ohr operon. The
bicistronic mRNA is then processed by RNase III-like enzymes to give
high levels of ohr mRNA, while ohrR mRNA
is rapidly degraded.
*
Corresponding author. Mailing address: Laboratory of
Biotechnology, Chulabhorn Research Institute, Lak Si, Bangkok 10210, Thailand. Phone: (662) 574-0623, ext. 1402. Fax: (662) 574-2027. E-mail: skorn{at}tubtim.cri.or.th.

Present address: School of Bioscience, University of Birmingham,
Edgbaston, Birmingham, United
Kingdom.
Journal of Bacteriology, August 2001, p. 4405-4412, Vol. 183, No. 15
0021-9193/01/$04.00+0 DOI: 10.1128/JB.183.15.4405-4412.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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