Bacillus subtilis cytochrome oxidase mutants: biochemical analysis and genetic evidence for two aa3-type oxidases.
The ctaBCDEF genes coding for cytochrome c oxidase were found to reside adjacent to a regulatory gene ctaA at 127 degrees on the Bacillus subtilis chromosome. The structural genes for subunits I and II, ctaD and ctaC, were deleted by gene-replacement using a phleomycin-resistance marker. The mutant was unable to oxidize N,N,N',N'-tetramethyl-p-phenylene-diamine and oxidized cytochrome c at a significantly lower rate. Absorption spectra of the mutant and wild-type membranes confirmed the presence of two haem A-containing enzymes in B. subtilis. Another mutant, with a spontaneous deletion upstream from ctaC, was found to express neither of these enzymes. Radioactive haem-labelling was used to identify subunit II, which contains a haem C, and cytochrome c-550 among the membrane-bound c-type cytochromes of B. subtilis.
European Molecular Biology Laboratory, Heidelberg, Germany.
J
J
van der Oost
C
C
von Wachenfeld
L
L
Hederstedt
M
M
Saraste
D001224:Aspartic Acid / Q000494:pharmacology; D001412:Bacillus subtilis / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D015139:Blotting, Southern; D002872:Chromosome Deletion; D003574:Cytochrome c Group / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017937:Cytochrome d Group; D003580:Cytochromes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004352:Drug Resistance, Microbial / Q000235:genetics; D003576:Electron Transport Complex IV / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005947:Glucose / Q000494:pharmacology; D005971:Glutamates / Q000494:pharmacology; D018698:Glutamic Acid; D009154:Mutation / Q000235:genetics; D010692:Phleomycins / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics; D013053:Spectrophotometry; D013772:Tetramethylphenylenediamine / Q000378:metabolism / Q000494:pharmacology
D003574:Cytochrome c Group; D003580:Cytochromes; D005971:Glutamates; D010692:Phleomycins; D001224:Aspartic Acid; D018698:Glutamic Acid; D017937:Cytochrome d Group; D003576:Electron Transport Complex IV; D005947:Glucose; D013772:Tetramethylphenylenediamine