Cloning, DNA sequence, and complementation analysis of the Salmonella typhimurium hemN gene encoding a putative oxygen-independent coproporphyrinogen III oxidase.
Coproporphyrinogen oxidation is a last step in heme biosynthesis. The biochemically characterized eukaryotic coproporphyrinogen III oxidases have an obligate requirement for molecular oxygen, and a similar enzyme is encoded by the hemF gene in Salmonella typhimurium. Anaerobic heme synthesis requires an oxygen-independent coproporphyrinogen oxidase, which is probably encoded by the hemN gene in S. typhimurium. The hemN gene has been cloned from an insertion mutant. The nucleotide sequence was obtained and used for PCR amplification of the wild-type gene. A single open reading frame was identified as the hemN gene on the basis of its interruption by the insertion mutation and plasmid complementation studies of hemF hemN double mutants. The predicted HemN protein has 38% amino acid sequence identity to a putative anaerobic Rhodobacter sphaeroides coproporphyrinogen oxidase. The hemN RNA 5' end and the inferred transcription initiation site were mapped by primer extension. The 52.8-kDa HemN protein is expressed from the second ATG codon of the hemN open reading frame. An open reading frame with an unknown function directly upstream of hemN has a striking amino acid sequence, including 11 acidic residues in a row.
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Department of Microbiology, University of Alabama at Birmingham 35294.
Elliott
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D000595:Amino Acid Sequence; D000693:Anaerobiosis; D001483:Base Sequence; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D003304:Coproporphyrinogen Oxidase / Q000235:genetics*; D003305:Coproporphyrinogens / Q000378:metabolism*; D005798:Genes, Bacterial / Q000235:genetics*; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D012486:Salmonella typhimurium / Q000201:enzymology / Q000235:genetics*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic
D003305:Coproporphyrinogens; C028026:coproporphyrinogen III; D003304:Coproporphyrinogen Oxidase