PMID:9022686

Citation	de Boer, AP, van der Oost, J, Reijnders, WN, Westerhoff, HV, Stouthamer, AH and van Spanning, RJ (1996) Mutational analysis of the nor gene cluster which encodes nitric-oxide reductase from Paracoccus denitrificans. Eur. J. Biochem. 242:592-600
Abstract	The genes that encode the hc-type nitric-oxide reductase from Paracoccus denitrificans have been identified. They are part of a cluster of six genes (norCBQDEF) and are found near the gene cluster that encodes the cd1-type nitrite reductase, which was identified earlier [de Boer, A. P. N., Reijnders, W. N. M., Kuenen, J. G., Stouthamer, A. H. & van Spanning, R. J. M. (1994) Isolation, sequencing and mutational analysis of a gene cluster involved in nitrite reduction in Paracoccus denitrificans, Antonie Leeu wenhoek 66, 111-127]. norC and norB encode the cytochrome-c-containing subunit II and cytochrome b-containing subunit I of nitric-oxide reductase (NO reductase), respectively. norQ encodes a protein with an ATP-binding motif and has high similarity to NirQ from Pseudomonas stutzeri and Pseudomonas aeruginosa and CbbQ from Pseudomonas hydrogenothermophila. norE encodes a protein with five putative transmembrane alpha-helices and has similarity to CoxIII, the third subunit of the aa3-type cytochrome-c oxidases. norF encodes a small protein with two putative transmembrane alpha-helices. Mutagenesis of norC, norB, norQ and norD resulted in cells unable to grow anaerobically. Nitrite reductase and NO reductase (with succinate or ascorbate as substrates) and nitrous oxide reductase (with succinate as substrate) activities were not detected in these mutant strains. Nitrite extrusion was detected in the medium, indicating that nitrate reductase was active. The norQ and norD mutant strains retained about 16% and 23% of the wild-type level of NorC, respectively. The norE and norF mutant strains had specific growth rates and NorC contents similar to those of the wild-type strain, but had reduced NOR and NIR activities, indicating that their gene products are involved in regulation of enzyme activity. Mutant strains containing the norCBQDEF region on the broad-host-range vector pEG400 were able to grow anaerobically, although at a lower specific growth rate and with lower NOR activity compared with the wild-type strain.
Links	PubMed
Keywords	Amino Acid Sequence; Base Sequence; Cloning, Molecular; DNA Mutational Analysis; Electron Transport Complex IV/chemistry; Genes, Bacterial; Genetic Complementation Test; Molecular Sequence Data; Nitric Oxide/metabolism; Operon; Oxidation-Reduction; Oxidoreductases/genetics; Paracoccus denitrificans/enzymology; Sequence Alignment; Sequence Homology, Amino Acid; Species Specificity
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
PARDE:NORC		GO:0016966: nitric oxide reductase activity	ECO:0000315:		F		Table 4.	complete CACAO 2491
PARDE:NORC	enables	GO:0016966: nitric oxide reductase activity	ECO:0000315: mutant phenotype evidence used in manual assertion		F		Seeded From UniProt	complete
PARDE:Q51665		GO:0016966: nitric oxide reductase activity	ECO:0000315:		F		Table 2	complete CACAO 2811
PARDE:Q51665	enables	GO:0016966: nitric oxide reductase activity	ECO:0000315: mutant phenotype evidence used in manual assertion		F		Seeded From UniProt	complete
PARDE:Q51666		GO:0016966: nitric oxide reductase activity	ECO:0000315:		F		Table 2	complete CACAO 2849
PARDE:Q51666	enables	GO:0016966: nitric oxide reductase activity	ECO:0000315: mutant phenotype evidence used in manual assertion		F		Seeded From UniProt	complete
PARDE:Q51667		GO:0016966: nitric oxide reductase activity	ECO:0000315:		F		Table 2	complete CACAO 2850
PARDE:Q51667	enables	GO:0016966: nitric oxide reductase activity	ECO:0000315: mutant phenotype evidence used in manual assertion		F		Seeded From UniProt	complete

References

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