Role of individual nap gene cluster products in NapC-independent nitrate respiration of Wolinella succinogenes
Abstract
Bacterial nap gene clusters, encoding periplasmic nitrate reductase (NapA), are complex and diverse, and the composition of the electron transport chain donating electrons to NapA is poorly characterized in most organisms. Exceptionally, Wolinella succinogenes transfers electrons from formate via the menaquinone pool to NapA independently of a membrane-bound c-type cytochrome of the NapC family. The role of individual ORFs of the W. succinogenes napAGHBFLD gene cluster is assessed here by characterizing in-frame gene inactivation mutants. The ability of the mutants to grow by nitrate respiration was tested and their NapA content and specific nitrate reductase activity were determined. The napB and napD gene products proved to be essential for nitrate respiration, with NapD being required for the production of mature NapA. Inactivation of either subunit of the putative membrane-bound menaquinol dehydrogenase complex NapGH almost abolished growth by nitrate respiration. Substitution of the twin-arginine sequence of NapG had the same effect as absence of NapG. Phenotypes of mutants lacking either NapF or NapL suggest that both proteins function in NapA assembly and/or export. The data substantiate the current model of the composition of the NapC-independent electron transport chain as well as of NapA maturation, and indicate the presence of an alternative electron transport pathway to NapA.
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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 [105]
NapB is an essential component of electron transport in organisms where it is present.
“This small c-type cytochrome is an essential component of electron transport in organisms where it is present.101,105”
From Nitrate and periplasmic nitrate reductases (2014) · cites this paper as [105]
NapF is not considered essential for NapA maturation, although it aids in [4Fe-4S] insertion.
“NapF is not considered to be essential for NapA maturation, although it does aid in [4Fe–4S] insertion. 105,258,274”
From Nitrate and periplasmic nitrate reductases (2014) · cites this paper as [105]
The gene napL encodes a 35 kDa periplasmic chaperone protein with no known metal binding motifs.
“The gene napL encodes a 35 kDa periplasmic chaperone protein,103,105 with no known metal binding motifs.103”
From Nitrate and periplasmic nitrate reductases (2014) · cites this paper as [105]
Mutation of napL in W. succinogenes and C. jejuni delays respiratory growth on nitrate media.
“While the function of NapL is unknown, mutation of napL in W. succinogenes and C. jejuni delays respiratory growth on nitrate media.105,276”
From Nitrate and periplasmic nitrate reductases (2014) · cites this paper as [105]
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- Authors
- Kern, M., Mager, A. M., & Simon, J.
- Affiliations
- Goethe University Frankfurt (DE)
- ORCID
- Simon, Jörg
- Venue
- Microbiology
- Publisher
- Microbiology Society
- Published
- 2007-11-01
- Volume
- 153
- Issue
- 11
- Pages
- 3739-3747
- ISSN
- 1350-0872 (ISSN-L)
- 1465-2080
- OpenAlex
- W2034189603
- Cited by
- 41
- References
- 42
- Citations by year
- 2026: 3 · 2025: 2 · 2024: 1 · 2023: 1 · 2020: 3 · 2019: 2 · 2018: 6 · 2017: 2 · 2016: 2 · 2015: 2 · 2014: 2 · 2013: 2 · 2012: 1
- FWCI
- 1.61
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Microbiology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- University of East Anglia
- Deutsche Forschungsgemeinschaft
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Physical Sciences › Environmental Science › Environmental Engineering › Microbial Fuel Cells and Bioremediation
- Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Metalloenzymes and iron-sulfur proteins
- Keywords
- Wolinella succinogenes, periplasmic nitrate reductase, napA, Napf, non-aqueous phase liquids, twin-arginine translocation, enzyme maturation, gene inactivation mutants
- MeSH terms
- Bacterial Proteins, Culture Media, Electron Transport, Multigene Family, Mutation, Nitrate Reductases, Nitrates, Gene Expression Regulation, Bacterial, Wolinella, Gene Deletion, Periplasm
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