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Cloning and characterization of the Bacillus subtilis hemEHY gene cluster, which encodes protoheme IX biosynthetic enzymes

Hansson, Mats LU and Hederstedt, Lars LU (1992) In Journal of Bacteriology 174(24). p.8081-8093
Abstract
Mutations that cause a block in a late step of the protoheme IX biosynthetic pathway, i.e., in a step after uroporphyrinogen III, map at 94 degrees on the Bacillus subtilis chromosomal genetic map. We have cloned and sequenced the hem genes at this location. The sequenced region contains six open reading frames: ponA, hemE, hemH, hemY, ORFA, and ORFB. The ponA gene product shows over 30% sequence identity to penicillin-binding proteins 1A of Escherichia coli, Streptococcus pneumoniae, and Streptococcus oralis and probably has a role in cell wall metabolism. The hemE gene was identified from amino acid sequence comparisons as encoding uroporphyrinogen III decarboxylase. The hemH gene was identified by enzyme activity analysis of the HemH... (More)
Mutations that cause a block in a late step of the protoheme IX biosynthetic pathway, i.e., in a step after uroporphyrinogen III, map at 94 degrees on the Bacillus subtilis chromosomal genetic map. We have cloned and sequenced the hem genes at this location. The sequenced region contains six open reading frames: ponA, hemE, hemH, hemY, ORFA, and ORFB. The ponA gene product shows over 30% sequence identity to penicillin-binding proteins 1A of Escherichia coli, Streptococcus pneumoniae, and Streptococcus oralis and probably has a role in cell wall metabolism. The hemE gene was identified from amino acid sequence comparisons as encoding uroporphyrinogen III decarboxylase. The hemH gene was identified by enzyme activity analysis of the HemH protein expressed in E. coli. It encodes a water-soluble ferrochelatase which catalyzes the final step in protoheme IX synthesis, the insertion of ferrous iron into protoporphyrin IX. The function of the hemY gene product was not elucidated, but mutation analysis shows that it is required for a late step in protoheme IX synthesis. The hemY gene probably encodes an enzyme with coproporphyrinogen III oxidase or protoporphyrinogen IX oxidase activity or both of these activities. Inactivation of the ORFA and ORFB genes did not block protoheme IX synthesis. Preliminary evidence for a hemEHY mRNA was obtained, and a promoter region located in front of hemE was identified. From these combined results we conclude that the hemEHY gene cluster encodes enzymes for the synthesis of protoheme IX from uroporphyrinogen III and probably constitutes an operon. (Less)
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published
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keywords
Mutation, *Muramoylpentapeptide Carboxypeptidase, *Multigene Family, Multienzyme Complexes/genetics, Molecular Sequence Data, Humans, Hexosyltransferases/genetics, Heme/biosynthesis/*genetics, Genetic Complementation Test, Ferrochelatase/genetics, Bacterial, DNA, Molecular, Cloning, *Carrier Proteins, Northern, Blotting, Base Sequence, *Bacterial Proteins, Bacillus subtilis/*enzymology/*genetics, Amino Acid Sequence, Animals, Open Reading Frames, Operon, Penicillin-Binding Proteins, Peptidyl Transferases/genetics, Restriction Mapping, Sequence Homology, Amino Acid, Uroporphyrinogen Decarboxylase/genetics
in
Journal of Bacteriology
volume
174
issue
24
pages
8081 - 8093
publisher
American Society for Microbiology
external identifiers
  • scopus:0026676696
ISSN
0021-9193
DOI
10.1128/jb.174.24.8081-8093.1992
language
English
LU publication?
yes
id
aec31e96-f4a8-49c1-a891-cbea1bf2a2de (old id 8001496)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/1459957
date added to LUP
2015-10-01 09:16:28
date last changed
2017-11-12 03:21:29
@article{aec31e96-f4a8-49c1-a891-cbea1bf2a2de,
  abstract     = {Mutations that cause a block in a late step of the protoheme IX biosynthetic pathway, i.e., in a step after uroporphyrinogen III, map at 94 degrees on the Bacillus subtilis chromosomal genetic map. We have cloned and sequenced the hem genes at this location. The sequenced region contains six open reading frames: ponA, hemE, hemH, hemY, ORFA, and ORFB. The ponA gene product shows over 30% sequence identity to penicillin-binding proteins 1A of Escherichia coli, Streptococcus pneumoniae, and Streptococcus oralis and probably has a role in cell wall metabolism. The hemE gene was identified from amino acid sequence comparisons as encoding uroporphyrinogen III decarboxylase. The hemH gene was identified by enzyme activity analysis of the HemH protein expressed in E. coli. It encodes a water-soluble ferrochelatase which catalyzes the final step in protoheme IX synthesis, the insertion of ferrous iron into protoporphyrin IX. The function of the hemY gene product was not elucidated, but mutation analysis shows that it is required for a late step in protoheme IX synthesis. The hemY gene probably encodes an enzyme with coproporphyrinogen III oxidase or protoporphyrinogen IX oxidase activity or both of these activities. Inactivation of the ORFA and ORFB genes did not block protoheme IX synthesis. Preliminary evidence for a hemEHY mRNA was obtained, and a promoter region located in front of hemE was identified. From these combined results we conclude that the hemEHY gene cluster encodes enzymes for the synthesis of protoheme IX from uroporphyrinogen III and probably constitutes an operon.},
  author       = {Hansson, Mats and Hederstedt, Lars},
  issn         = {0021-9193},
  keyword      = {Mutation,*Muramoylpentapeptide Carboxypeptidase,*Multigene Family,Multienzyme Complexes/genetics,Molecular Sequence Data,Humans,Hexosyltransferases/genetics,Heme/biosynthesis/*genetics,Genetic Complementation Test,Ferrochelatase/genetics,Bacterial,DNA,Molecular,Cloning,*Carrier Proteins,Northern,Blotting,Base Sequence,*Bacterial Proteins,Bacillus subtilis/*enzymology/*genetics,Amino Acid Sequence,Animals,Open Reading Frames,Operon,Penicillin-Binding Proteins,Peptidyl Transferases/genetics,Restriction Mapping,Sequence Homology,Amino Acid,Uroporphyrinogen Decarboxylase/genetics},
  language     = {eng},
  number       = {24},
  pages        = {8081--8093},
  publisher    = {American Society for Microbiology},
  series       = {Journal of Bacteriology},
  title        = {Cloning and characterization of the <em>Bacillus subtilis hemEHY</em> gene cluster, which encodes protoheme IX biosynthetic enzymes},
  url          = {http://dx.doi.org/10.1128/jb.174.24.8081-8093.1992},
  volume       = {174},
  year         = {1992},
}