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Isolation and Characterization of rpoS from a Pathogenic Bacterium, Vibrio vulnificus: Role of ^S in Survival of Exponential-Phase Cells under Oxidative Stress

Kyung-Je Park,¹ Min-Jin Kang,^1, Songhee H. Kim,² Hyun-Jung Lee,¹ Jae-Kyu Lim,¹ Sang Ho Choi,³ Soon-Jung Park,⁴ and Kyu-Ho Lee^1*

Department of Environmental Science, Hankuk University of Foreign Studies, Yongin, Kyunggi-Do 449-791,¹ Department of Food Science and Technology, Seoul National University, Seoul 151-742,³ Department of Parasitology, Yonsei University College of Medicine, Seoul 133-791, South Korea,⁴ Department of Cell Biology and Molecular Genetics, University of Maryland at College Park, College Park, Maryland²

Received 3 September 2003/ Accepted 24 February 2004

A gene homologous to rpoS was cloned from a fatal human pathogen, Vibrio vulnificus. The functional role of rpoS in V. vulnificus was accessed by using an rpoS knockout mutant strain. This mutant was impaired in terms of the ability to survive under oxidative stress, nutrient starvation, UV irradiation, or acidic conditions. The increased susceptibility of the V. vulnificus mutant in the exponential phase to H₂O₂ was attributed to the reduced activity of hydroperoxidase I (HPI). Although ^S synthesis was induced and HPI activity reached the maximal level in the stationary phase, the mutant in the stationary phase showed the same susceptibility to H₂O₂ as the wild-type strain in the stationary phase. In addition, HPII activity, which is known to be controlled by ^S in Escherichia coli, was not detectable in V. vulnificus strains under the conditions tested. The mutant in the exponential phase complemented with multiple copies of either the rpoS or katG gene of V. vulnificus recovered both resistance to H₂O₂ and HPI activity compared with the control strain. Expression of the katG gene encoding HPI in V. vulnificus was monitored by using a katG::luxAB transcriptional fusion. The expression of this gene was significantly reduced by deletion of ^S in both the early exponential and late stationary phases. Thus, ^S is necessary for increased synthesis and activity of HPI, and ^S is required for exponentially growing V. vulnificus to develop the ability to survive in the presence of H₂O₂.

Present address: Department of Microbiology, University of Idaho, Moscow, Idaho.

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