PMID:9079911

Citation	Kerby, RL, Ludden, PW and Roberts, GP (1997) In vivo nickel insertion into the carbon monoxide dehydrogenase of Rhodospirillum rubrum: molecular and physiological characterization of cooCTJ. J. Bacteriol. 179:2259-66
Abstract	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)omega-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 microM Ni supplementation, strains bearing Gm(r)omega-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 microM Ni.
Links	PubMed PMC178962
Keywords	Aldehyde Oxidoreductases/chemistry; Aldehyde Oxidoreductases/physiology; Amino Acid Sequence; Bacterial Proteins/genetics; Binding Sites; Carrier Proteins/genetics; Carrier Proteins/physiology; Electrophoresis, Gel, Two-Dimensional; Escherichia coli Proteins; Membrane Proteins; Molecular Sequence Data; Multienzyme Complexes/chemistry; Multienzyme Complexes/physiology; Nickel/metabolism; Operon; Recombinant Proteins; Rhodospirillum rubrum/enzymology; Rhodospirillum rubrum/genetics; Structure-Activity Relationship
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
RHORU:COOC		GO:0018414: nickel incorporation into metallo-sulfur cluster	ECO:0000315: mutant phenotype evidence used in manual assertion		P		Figure 4 shows Ni concentration dependence of photosynthetic and CO-dependent growth of R. rubrum strains. Strains were mutated resulting in the gene cooFSCTJA being deleted.	complete
RHORU:COOC	involved_in	GO:0018414: nickel incorporation into metallo-sulfur cluster	ECO:0000315: mutant phenotype evidence used in manual assertion		P		Seeded From UniProt	complete

References

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