Nickel-deficient carbon monoxide dehydrogenase from Rhodospirillum rubrum: in vivo and in vitro activation by exogenous nickel.

Bonam, Duane; McKenna, M. C.; Stephens, Philip J.; Ludden, Paul W.

doi:10.1073/pnas.85.1.31

Cited by 58 publications

(75 citation statements)

References 19 publications

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“…Furthermore, insertion of a cassette immediately upstream from the cooHencoded C-terminal conserved region generated a mutant that contained active CODH yet failed to produce H2 in the presence of CO, again consistent with the identification of cooH. However, comparing the R. rubrum CO-induced hydrogenase with other Ni hydrogenases may not be appropriate, as the presence of Ni in this enzyme is not proved (though its function depends on Ni in the medium [12]), and the effect of the insertion is consistent with cooH encoding another protein involved in electron transfer from CODH to the hydrogenase. Notably, CooH shows considerable similarity to an NADH-ubiquinone oxidoreductase (23).…”

Section: Discussionmentioning

confidence: 96%

Genetic and physiological characterization of the Rhodospirillum rubrum carbon monoxide dehydrogenase system

et al. 1992

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(38,61). The enzymes generally have high affinity for CO and couple its oxidation in vitro with the reduction of acceptors such as methylene blue; most do not reduce low-potential acceptors. The purified enzymes contain a molybdenum cofactor plus Fe and perhaps Se, Zn, and Cu (38). The structural genes from Pseudomonas thermocarboxydovorans have been cloned (5).In anaerobic organisms, including acetogens, dissimilatory sulfate reducers, and methanogens, the CODHs are constitutively synthesized metalloproteins, usually 02 labile, that catalyze the interconversion of single-carbon moieties with acetyl coenzyme A (acetyl-CoA); the readily assayed oxidation of CO to CO2 (coupled with the reduction of low-potential electron acceptors) is but one catalytic function. The interconversion is a fundamental anaerobic metabolic process, and the physiology and biochemistry of these bacteria have been the subjects of several reviews (29,33,43,54,62

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Section: Discussionmentioning

confidence: 96%

Genetic and physiological characterization of the Rhodospirillum rubrum carbon monoxide dehydrogenase system

et al. 1992

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“…It is a reasonable hypothesis that a metal serves as a CO-binding site in CooA, since CO interacts with proteins by binding to metals or metal centers in virtually all described cases. This is true for proteins that utilize CO as a substrate, such as the CO dehydrogenases from R. rubrum (6,13,14) and Clostridium thermoaceticum (31,41,46). CO also interacts with metals in proteins that are inhibited by CO, including hydrogenases from Clostridium pasteurianum (1) and Chromatium vinosum (50).…”

Section: Discussionmentioning

confidence: 99%

“…The system consists of at least three proteins that together couple CO oxidation to H 2 evolution: CooS (CO dehydrogenase), which oxidizes CO; CooF, a CooS-associated Fe-S protein; and CooH, a CO-tolerant hydrogenase (4,5,15). CooS and CooF have been purified to homogeneity and biochemically characterized (5,6,(13)(14)(15). The cooH, cooF, and cooS genes are adjacent to one another on the chromosome and have been cloned, sequenced, and mutationally characterized.…”

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Carbon monoxide-induced activation of gene expression in Rhodospirillum rubrum requires the product of cooA, a member of the cyclic AMP receptor protein family of transcriptional regulators

et al. 1995

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Induction of the CO-oxidizing system of the photosynthetic bacterium Rhodospirillum rubrum is regulated at the level of gene expression by the presence of CO. In this paper, we describe the identification of a gene that is required for CO-induced gene expression. An 11-kb deletion of the region adjacent to the previously characterized cooFSCTJ region resulted in a mutant unable to synthesize CO dehydrogenase in response to CO and unable to grow utilizing CO as an energy source. A 2.5-kb region that corresponded to a portion of the deleted region complemented this mutant for its CO regulation defect, restoring its ability to grow utilizing CO as an energy source. When the 2.5-kb region was sequenced, one open reading frame, designated cooA, predicted a product showing similarity to members of the cyclic AMP receptor protein (CRP) family of transcriptional regulators. The product, CooA, is 28% identical (51% similar) to CRP and 18% identical (45% similar) to FNR from Escherichia coli. The insertion of a drug resistance cassette into cooA resulted in a mutant that could not grow utilizing CO as an energy source. CooA contains a number of cysteine residues substituted at, or adjacent to, positions that correspond to residues that contact cyclic AMP in the crystal structure of CRP. A model based on this observation is proposed for the recognition of CO by CooA. Adjacent to cooA are two genes, nadB and nadC, with predicted products similar to proteins in other bacteria that catalyze reactions in the de novo synthesis of NAD. A mutant with the cooA-nadBC region deleted displayed an auxotrophy for nicotinic acid, while the cooA insertion mutant did not.The purple, nonsulfur, phototrophic bacterium Rhodospirillum rubrum synthesizes an enzyme system for carbon monoxide (CO) oxidation. Anaerobically in the dark, R. rubrum can utilize the CO-oxidizing system to generate energy (28). The system consists of at least three proteins that together couple CO oxidation to H 2 evolution: CooS (CO dehydrogenase), which oxidizes CO; CooF, a CooS-associated Fe-S protein; and CooH, a CO-tolerant hydrogenase (4,5,15). CooS and CooF have been purified to homogeneity and biochemically characterized (5, 6, 13-15). The cooH, cooF, and cooS genes are adjacent to one another on the chromosome and have been cloned, sequenced, and mutationally characterized. Mutations in any of these genes eliminate the ability of R. rubrum to evolve H 2 from CO and to utilize CO as an energy source (27,28).Unlike the case for many other bacteria capable of oxidizing CO anaerobically (11), the presence of the CO-oxidizing system in R. rubrum is dependent upon exogenous CO. Induction of this system by CO occurs in cells grown either photoheterotrophically (4, 5) or anaerobically in the dark (28). For cells growing photoheterotrophically in malate-ammonium medium, CooS activity is induced at least 1,000-fold by CO; under these conditions, CooS can constitute 2 to 5% of cellular protein (5). CooS accumulation following exposure to CO requires protein synthesis, in...

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“…Limited evidence suggests that accessory functions are necessary for posttranslational Ni insertion and CODH activity in R. rubrum: an Ni-deficient apo-CODH is found in induced cultures deprived of Ni; substantially higher Ni concentrations are required for activation of apo-CODH in vitro than in vivo; and there is an elevated Ni requirement for CO-dependent growth of a mutant bearing an insertion in cooC, a gene immediately downstream of the CODH structural gene (5,13,31).…”

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In vivo nickel insertion into the carbon monoxide dehydrogenase of Rhodospirillum rubrum: molecular and physiological characterization of cooCTJ

1997

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The products of cooCTJ are involved in normal in vivo Ni insertion into the carbon monoxide dehydrogenase (CODH) of Rhodospirillum rubrum. Located on a 1.5-kb DNA segment immediately downstream of the CODH structural gene (cooS), two of the genes encode proteins that bear motifs reminiscent of other (urease and hydrogenase) Ni-insertion systems: a nucleoside triphosphate-binding motif near the N terminus of CooC and a run of 15 histidine residues regularly spaced over the last 30 amino acids of the C terminus of CooJ. A Gm r ⍀-linker cassette was developed to create both polar and nonpolar (60 bp) insertions in the cooCTJ region, and these, along with several deletions, were introduced into R. rubrum by homologous recombination. Analysis of the exogenous Ni levels required to sustain CO-dependent growth of the R. rubrum mutants demonstrated different phenotypes: whereas the wild-type strain and a mutant bearing a partial cooJ deletion (of the region encoding the histidine-rich segment) grew at 0.5 M Ni supplementation, strains bearing Gm r ⍀-linker cassettes in cooT and cooJ required approximately 50-fold-higher Ni levels and all cooC insertion strains, bearing polar or nonpolar insertions, grew optimally at 550 M Ni.The phototrophic bacterium Rhodospirillum rubrum induces synthesis of an Ni-and Fe-containing carbon monoxide dehydrogenase (CODH) upon anaerobic exposure to CO (3, 4) and couples CO oxidation to H 2 evolution as a source of energy (31). Limited evidence suggests that accessory functions are necessary for posttranslational Ni insertion and CODH activity in R. rubrum: an Ni-deficient apo-CODH is found in induced cultures deprived of Ni; substantially higher Ni concentrations are required for activation of apo-CODH in vitro than in vivo; and there is an elevated Ni requirement for CO-dependent growth of a mutant bearing an insertion in cooC, a gene immediately downstream of the CODH structural gene (5,13,31).Aside from the physiology of Ni insertion, the R. rubrum CODH has been well characterized biochemically (4, 14), genes encoding its synthesis and several additional proteins likely involved in H 2 production have been cloned and sequenced (16,17,30), active CODH has been heterologously expressed (24), and the CO-responsive transcriptional regulator is under investigation (24, 53). The hydrogenase (CooH) and CODH (CooS) genes occur in two operons, cooMKLXUH and cooFSCTJ, whose coding regions are separated by a 450-bp interval, with the CO-responsive transcriptional activator (CooA) encoded 137 nucleotides downstream of cooJ.Formation of the Ni-containing center(s) of other Ni-CODH enzymes has not been characterized, although the activities are prevalent in diverse anaerobic archaea and bacteria (15). Both the spectroscopic similarities of purified enzymes (26) and evident conservation of a limited number of potential metal ligands in an alignment of bacterial and archaeal CODH sequences (e.g., see reference 30) are consistent with conservation of structure and function of the Ni-containing C center...

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Nickel-deficient carbon monoxide dehydrogenase from Rhodospirillum rubrum: in vivo and in vitro activation by exogenous nickel.

Cited by 58 publications

References 19 publications

Genetic and physiological characterization of the Rhodospirillum rubrum carbon monoxide dehydrogenase system

Genetic and physiological characterization of the Rhodospirillum rubrum carbon monoxide dehydrogenase system

Carbon monoxide-induced activation of gene expression in Rhodospirillum rubrum requires the product of cooA, a member of the cyclic AMP receptor protein family of transcriptional regulators

In vivo nickel insertion into the carbon monoxide dehydrogenase of Rhodospirillum rubrum: molecular and physiological characterization of cooCTJ

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