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Eur J Biochem ; 268(8): 2486-97, 2001 Apr.
Article in English | MEDLINE | ID: mdl-11298768

ABSTRACT

In order to study the induction of terminal oxidases in Paracoccus denitrificans, their promoters were fused to the lacZ reporter gene and analysed in the wild-type strain, in an FnrP-negative mutant, in a cytochrome bc1-negative mutant, and in six single or double oxidase-negative mutant strains. The strains were grown under aerobic, semi-aerobic, and denitrifying conditions. The oxygen-sensing transcriptional-regulatory protein FnrP negatively regulated the activity of the qox promoter, which controls expression of the ba3-type quinol oxidase, while it positively regulated the activity of the cco promoter, which controls expression of the cbb3-type cytochrome c oxidase. The ctaDII and ctaC promoters, which control the expression of the aa3-type cytochrome c oxidase subunits I and II, respectively, were not regulated by FnrP. The activities of the latter two promoters, however, did decrease with decreasing oxygen concentrations in the growth medium, suggesting that an additional oxygen-sensing mechanism exists that regulates transcription of ctaDII and ctaC. Apparently, the intracellular oxygen concentration (as sensed by FnrP) was not the only signal to which the oxidase promoters responded. At given extracellular oxygen status, both the qox and the cco promoters responded to mutations in terminal oxidase genes, whereas the ctaDII and ctaC promoters did not. The change of electron distribution through the respiratory network, resulting from elimination of one or more oxidase genes, may have changed intracellular signals that affect the activities of the qox and cco promoters. On the other hand, the re-routing of electron distribution in the respiratory mutants hardly affected the oxygen consumption rate as compared to that of the wild-type. This suggests that the mutants adapted their respiratory network in such a way that they were able to consume oxygen at a rate similar to that of the wild-type strain.


Subject(s)
Gene Expression Regulation, Enzymologic , Oxidoreductases/chemistry , Paracoccus denitrificans/enzymology , Base Sequence , Cell Membrane/metabolism , Cloning, Molecular , Cytochrome c Group/metabolism , Electron Transport Complex IV/metabolism , Electrons , Electrophoresis, Polyacrylamide Gel , Escherichia coli/metabolism , Lac Operon , Molecular Sequence Data , Mutagenesis, Site-Directed , Mutation , Oxidoreductases/genetics , Oxygen/metabolism , Oxygen Consumption , Plasmids/metabolism , Promoter Regions, Genetic , Sequence Homology, Nucleic Acid , Time Factors , beta-Galactosidase/metabolism
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