Cited by 56Bacterial Genetics and BiotechnologyMetalloenzymes and iron-sulfur proteinsWastewater Treatment and Nitrogen Removal

Periplasmic nitrate-reducing system of the phototrophic bacterium Rhodobacter sphaeroides DSM 158: transcriptional and mutational analysis of the napKEFDABC gene cluster

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REYES, Francisca, GAVIRA, Mónica, CASTILLO, Francisco, MORENO-VIVIÁN, Conrado·1998·Biochemical Journal

Abstract

The phototrophic bacterium Rhodobacter sphaeroidesDSM 158 is able to reduce nitrate to nitrite by means of a periplasmic nitrate reductase which is induced by nitrate and is not repressed by ammonium or oxygen. Recently, a 6.8 kb PstI DNA fragment carrying the napABCgenes coding for this periplasmic nitrate-reducing system was cloned [Reyes, Roldán, Klipp, Castillo and Moreno-Vivián (1996) Mol. Microbiol. 19, 1307–1318]. Further sequence and genetic analyses of the DNA region upstream from the napABCgenes reveal the presence of four additional napgenes. All these R. sphaeroidesgenes seem to be organized into a napKEFDABCtranscriptional unit. In addition, a partial open reading frame similar to the Azorhizobium caulinodans yntCgene and the Escherichia coli yjcCand yhjKgenes is present upstream from this napgene cluster. The R. sphaeroides napKgene codes for a putative 6.3 kDa transmembrane protein which is not similar to known proteins and the napEgene codes for a 6.7 kDa transmembrane protein similar to the Thiosphaera pantotrophaNapE. The R. sphaeroides napFgene product is a 16.4 kDa protein with four cysteine clusters that probably bind four [4Fe-4S] centres. This iron–sulphur protein shows similarity to the NapF and NapG proteins of E. coliand Haemophilus influenzae.Finally, the napDgene product is a 9.4 kDa soluble protein which is also found in E. coliand T. pantotropha. The 5´ end of the naptranscript has been determined by primer extension, and a δ70-like promoter has been identified upstream from the napKgene. The same transcriptional start site is found for cells growing aerobically or anaerobically with nitrate. Different mutant strains carrying defined polar and non-polar insertions in each napgene were constructed. Characterization of these mutant strains demonstrates the participation of the napgene products in the periplasmic nitrate reduction in R. sphaeroides.

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  • napB gene deletion mutants in R. sphaeroides and E. coli exhibit very little activity.

    “napB gene deletion mutants in R. sphaeroides215 and E. coli102 exhibit very little activity”

    From Nitrate and periplasmic nitrate reductases (2014) · cites this paper as [215]

  • Mutational studies on R. sphaeroides, E. coli, and Wolinella succinogenes have given insight into the significance of each auxiliary nap gene in nitrate reduction via Nap.

    “Mutational studies conducted on R. sphaeroides,215 E. coli,102 and Wolinella succinogenes105 have given insight into the significance of each auxiliary nap gene in nitrate reduction via Nap.”

    From Nitrate and periplasmic nitrate reductases (2014) · cites this paper as [215]

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  1. Sanger, F., Nicklen, S. and Coulson, A. R. (1977) Proc. Natl. Acad. Sci. U.S.A. 74, 5463-5467

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Authors
REYES, F., GAVIRA, M., CASTILLO, F., & MORENO-VIVIÁN, C.
Affiliations
  • University of Córdoba (ES)
  • Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de Córdoba, 14071-Córdoba, Spain
Venue
Biochemical Journal
Published
1998-05-01
Volume
331
Issue
3
Pages
897-904
ISSN
  • 0264-6021 (ISSN-L)
  • 1470-8728
OpenAlex
W2139697583
Cited by
56
References
44
Citations by year
2026: 1 · 2023: 3 · 2020: 1 · 2019: 1 · 2018: 5 · 2015: 1 · 2014: 3 · 2013: 2 · 2012: 1
FWCI
3.25
Type
journal article
Language
en
Open access
Locations
  • Biochemical Journal (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • Universidad de Sevilla
  • University of East Anglia
  • University of Oxford
  • Universidad de Córdoba
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Metalloenzymes and iron-sulfur proteins
  • Physical Sciences › Environmental Science › Pollution › Wastewater Treatment and Nitrogen Removal
Keywords
Rhodobacter sphaeroides, periplasmic nitrate reductase, nitrate reduction, nitrite, sigma 70 promoter, transcription start sites, Thiosphaera pantotropha
MeSH terms
Amino Acid Sequence, Bacterial Proteins, Cloning, Molecular, DNA Mutational Analysis, Electron Transport, Genes, Bacterial, Molecular Sequence Data, Nitrate Reductases, Paraquat, Promoter Regions, Genetic, Rhodobacter sphaeroides, Transcription, Genetic, Restriction Mapping, Gene Expression Regulation, Enzymologic, Mutagenesis, Insertional, Sequence Homology, Amino Acid, Sequence Analysis, DNA, Periplasm, Nitrate Reductase

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