Isolation and properties of reduced nicotinamide adenine dinucleotide-rubredoxin oxidoreductase of Clostridium acetobutylicum

Petitdemange, H.; Marczak, R.; Blusson, Hubert; Gay, Robert

doi:10.1016/0006-291x(79)91202-6

Cited by 41 publications

(20 citation statements)

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“…These essentially include oxygen-reducing flavodiiron proteins (FDPs), superoxide-reducing desulfoferrodoxin (Dfx), and rubrerythrins as peroxidases (26,56,57). Reduced rubredoxin thereby acts as the intermediate electron carrier, the pool of which is continuously reloaded by an NADH-dependent rubredoxin-dependent oxidoreductase (55,57). The sporulating gram-positive anaerobe Clostridium acetobutylicum is a classic example of a fermentative strict anaerobe.…”

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The Role of PerR in O₂-Affected Gene Expression ofClostridium acetobutylicum

et al. 2009

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In the strict anaerobe Clostridium acetobutylicum, a PerR-homologous protein has recently been identified as being a key repressor of a reductive machinery for the scavenging of reactive oxygen species and molecular O 2 . In the absence of PerR, the full derepression of its regulon resulted in increased resistance to oxidative stress and nearly full tolerance of an aerobic environment. In the present study, the complementation of a Bacillus subtilis PerR mutant confirmed that the homologous protein from C. acetobutylicum acts as a functional peroxide sensor in vivo. Furthermore, we used a transcriptomic approach to analyze gene expression in the aerotolerant PerR mutant strain and compared it to the O 2 stimulon of wild-type C. acetobutylicum. The genes encoding the components of the alternative detoxification system were PerR regulated. Only few other targets of direct PerR regulation were identified, including two highly expressed genes encoding enzymes that are putatively involved in the central energy metabolism. All of them were highly induced when wild-type cells were exposed to sublethal levels of O 2 . Under these conditions, C. acetobutylicum also activated the repair and biogenesis of DNA and Fe-S clusters as well as the transcription of a gene encoding an unknown CO dehydrogenase-like enzyme. Surprisingly few genes were downregulated when exposed to O 2 , including those involved in butyrate formation. In summary, these results show that the defense of this strict anaerobe against oxidative stress is robust and by far not limited to the removal of O 2 and its reactive derivatives.

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The Role of PerR in O₂-Affected Gene Expression ofClostridium acetobutylicum

et al. 2009

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“…Several attempts have been made to define the redox partners of rubredoxin by biochemical experiments. By using the rubredoxin-mediated reduction of eucaryotic cytochrome c by NADH as an assay, an NADH-rubredoxin oxidoreductase was (partially) purified from D. gigas (16) and C. acetobutylicum (25). The Clostridium enzyme was found to have flavin adenine dinucleotide as the prosthetic group and to consist of a single subunit with a molecular mass of 41 kDa (25) A different approach to the identification of possible redox * Corresponding author.…”

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Analysis of the transcriptional unit encoding the genes for rubredoxin (rub) and a putative rubredoxin oxidoreductase (rbo) in Desulfovibrio vulgaris Hildenborough

Brumlik

Voordouw

1989

J Bacteriol

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The nucleotide sequence of a 2.0-kilobase-pair EcoRI restriction fragment upstream from the gene (rub, 162 base pairs) encoding rubredoxin from Desulfovibrio vulgaris Hildenborough indicates that it is part of a larger transcriptional unit, containing an additional 378-base-pair open reading frame which terminates 16 nucleotides from the translational start of the rub gene and could encode a polypeptide of 14 kilodaltons (kDa). Northern (RNA) blotting of RNA isolated from both D. vulgaris Hildenborough and Escherichia coli TG2 transformed with plasmid pJK29, which contains both genes on a 1.1-kilobase-pair SalI insert, confirms that the genes for this 14-kDa polypeptide and rubredoxin are present on a single transcript of 680 nucleotides. Strong evidence that the 14-kDa polypeptide is also a redox protein is provided by the fact that its NH2 terminus is homologous to desulforedoxin, which has been isolated from D. gigas as a small dimeric redox protein (36 amino acids per monomer), coordinating two iron atoms. Since rubredoxin is a potential redox partner for the 14-kDa protein, it has been tentatively named rubredoxin oxidoreductase, produced by the rbo gene. Southern blotting indicates that the rbo-rub operon is present in several species and strains of sulfate-reducing bacteria.

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“…S1B in the supplemental material). NROR has previously been reported to function as an NADH-dependent Rd reductase in vitro (29). However, the in vivo function of this enzyme has not yet been elucidated (9,29).…”

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“…NROR has previously been reported to function as an NADH-dependent Rd reductase in vitro (29). However, the in vivo function of this enzyme has not yet been elucidated (9,29). It has also been reported that reactivity of NROR toward O 2 could not be detected (29).…”

Section: Resultsmentioning

confidence: 99%

“…A H 2 O-forming NADH oxidase homologue is not present in the C. acetobutylicum genome, though NROR does have a low homology (17% identity) to this enzyme (18). The C. acetobutylicum NROR was originally characterized as an NADH:rubredoxin oxidoreductase, although a function of NROR in protection against oxidative stress has not been elucidated (9,29). Although we have shown that nror operon genes are responsive to O 2 stress, the NROR enzyme lacks a cysteine residue at the active center, which implies that NROR lacks an NADH-dependent O 2 reductase activity (18).…”

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O ₂ and Reactive Oxygen Species Detoxification Complex, Composed of O ₂ -Responsive NADH:Rubredoxin Oxidoreductase-Flavoprotein A2-Desulfoferrodoxin Operon Enzymes, Rubperoxin, and Rubredoxin, in Clostridium acetobutylicum

Kawasaki

Sakai

Takahashi

et al. 2009

Appl Environ Microbiol

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Isolation and properties of reduced nicotinamide adenine dinucleotide-rubredoxin oxidoreductase of Clostridium acetobutylicum

Cited by 41 publications

References 15 publications

The Role of PerR in O₂-Affected Gene Expression ofClostridium acetobutylicum

The Role of PerR in O₂-Affected Gene Expression ofClostridium acetobutylicum

Analysis of the transcriptional unit encoding the genes for rubredoxin (rub) and a putative rubredoxin oxidoreductase (rbo) in Desulfovibrio vulgaris Hildenborough

O ₂ and Reactive Oxygen Species Detoxification Complex, Composed of O ₂ -Responsive NADH:Rubredoxin Oxidoreductase-Flavoprotein A2-Desulfoferrodoxin Operon Enzymes, Rubperoxin, and Rubredoxin, in Clostridium acetobutylicum

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Isolation and properties of reduced nicotinamide adenine dinucleotide-rubredoxin oxidoreductase of Clostridium acetobutylicum

Cited by 41 publications

References 15 publications

The Role of PerR in O2-Affected Gene Expression ofClostridium acetobutylicum

The Role of PerR in O2-Affected Gene Expression ofClostridium acetobutylicum

Analysis of the transcriptional unit encoding the genes for rubredoxin (rub) and a putative rubredoxin oxidoreductase (rbo) in Desulfovibrio vulgaris Hildenborough

O 2 and Reactive Oxygen Species Detoxification Complex, Composed of O 2 -Responsive NADH:Rubredoxin Oxidoreductase-Flavoprotein A2-Desulfoferrodoxin Operon Enzymes, Rubperoxin, and Rubredoxin, in Clostridium acetobutylicum

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The Role of PerR in O₂-Affected Gene Expression ofClostridium acetobutylicum

The Role of PerR in O₂-Affected Gene Expression ofClostridium acetobutylicum

O ₂ and Reactive Oxygen Species Detoxification Complex, Composed of O ₂ -Responsive NADH:Rubredoxin Oxidoreductase-Flavoprotein A2-Desulfoferrodoxin Operon Enzymes, Rubperoxin, and Rubredoxin, in Clostridium acetobutylicum