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Analysis of theRhodobacter sphaeroides 2.4.3 genome revealed four previously unidentified sequences similar to the binding site of the transcriptional regulator NnrR. Expression studies demonstrated that three of these sequences are within the promoters of genes, designatedpaz ,norEF , andcdgA , in the NnrR regulon, while the status of the fourth sequence, within thetat operon promoter, remains uncertain.nnrV , under control of a previously identified NnrR site, was also identified.paz encodes a pseudoazurin that is a donor of electrons to nitrite reductase.paz inactivation did not decrease nitrite reductase activity, but loss of pseudoazurin and cytochromec2 together reduced nitrite reduction. Inactivation ofnorEF reduced nitrite and nitric oxide reductase activity and increased the sensitivity to nitrite in a taxis assay. This suggests that loss ofnorEF increases NO production as a result of decreased nitric oxide reductase activity. 2.4.3 is the only strain ofR. sphaeroides withnorEF , even though all four of the strains whose genomes have been sequenced have thenorCBQD operon andnnrR .norEF was shown to provide resistance to nitrite when it was mobilized intoR. sphaeroides strain 2.4.1 containingnirK . Inactivation of the other identified genes did not reveal any detectable denitrification-related phenotype. The distribution of members of the NnrR regulon inR. sphaeroides revealed patterns of coselection of structural genes with the ancillary genes identified here. The strong coselection of these genes indicates their functional importance under real-world conditions, even though inactivation of the majority of them does not impact denitrification under laboratory conditions.

Identification, Functional Studies, and Genomic Comparisons of New Members of the NnrR Regulon in Rhodobacter sphaeroides