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Ohr , Encoding An Organic Hydroperoxide Reductase, Is An In Vivo-Induced Gene In Actinobacillus Pleuropneumoniae

Robin J. Shea, Martha H. Mulks

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ABSTRACT Actinobacillus pleuropneumoniae is the causative agent of porcine pleuropneumonia, a disease characterized by pulmonary necrosis and hemorrhage caused in part by neutrophil degranulation. In an effort to understand the pathogenesis of this disease, we have developed an in vivo expression technology (IVET) system to identify genes that are specifically up-regulated during infection. One of the genes that we have identified as being induced in vivo is ohr , encoding organic hydroperoxide reductase, an enzyme that could play a role in detoxification of organic hydroperoxides generated during infection. Among the 12 serotypes of A . pleuropneumoniae , ohr was found in only serotypes 1, 9, and 11. This distribution correlated with increased resistance to cumene hydroperoxide, an organic hydroperoxide, but not to hydrogen peroxide or to paraquat, a superoxide generator. Functional assays of Ohr activity demonstrated that A . pleuropneumoniae serotype 1 cultures, but not serotype 5 cultures, were able to degrade cumene hydroperoxide. In A . pleuropneumoniae serotype 1, expression of ohr was induced by cumene hydroperoxide, but not by either hydrogen peroxide or paraquat. In contrast, an ohr gene from serotype 1 cloned into A . pleuropneumoniae serotype 5 was not induced by cumene hydroperoxide or by other forms of oxidative stress, suggesting the presence of a serotype-specific positive regulator of ohr in A . pleuropneumoniae serotype 1.