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Heme utilization in Campylobacter jejuni

By Kristian A. Ridley, Jonathan D. Rock, Ying Li and Julian M. Ketley

Abstract

A putative iron- and Fur-regulated hemin uptake gene cluster, composed of the transport genes chuABCD and a putative heme oxygenase gene (Cj1613c), has been identified in Campylobacter jejuni NCTC 11168. Mutation of chuA or Cj1613c leads to an inability to grow in the presence of hemin or hemoglobin as a sole source of iron. Mutation of chuB, -C, or -D only partially attenuates growth where hemin is the sole iron source, suggesting that an additional inner membrane (IM) ABC (ATP-binding cassette) transport system(s) for heme is present in C. jejuni. Genotyping experiments revealed that Cj1613c is highly conserved in 32 clinical isolates. One strain did not possess chuC, though it was still capable of using hemin/hemoglobin as a sole iron source, supporting the hypothesis that additional IM transport genes are present. In two other strains, sequence variations within the gene cluster were apparent and may account for an observed negative heme utilization phenotype. Analysis of promoter activity within the Cj1613c-chuA intergenic spacer region revealed chuABCD and Cj1613c are expressed from separate iron-repressed promoters and that this region also specifically binds purified recombinant FurCj in gel retardation studies. Absorbance spectroscopy of purified recombinant His6-Cj1613c revealed a 1:1 heme:His6-Cj1613c binding ratio. The complex was oxidatively degraded in the presence of ascorbic acid as the electron donor, indicating that the Cj1613c gene product functions as a heme oxygenase. In conclusion, we confirm the involvement of Cj1613c and ChuABCD in heme/hemoglobin utilization in C. jejuni.Peer-reviewedPublisher Versio

Publisher: American Society for Microbiology
Year: 2006
DOI identifier: 10.1128/JB.00994-06
OAI identifier: oai:lra.le.ac.uk:2381/2055
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  1. (1995). A gene cluster involved in the utilization of both free heme and heme:hemopexin by Haemophilus influenzae type b.
  2. (1997). A new type of hemophoredependent heme acquisition system of Serratia marcescens reconstituted in Escherichia coli.
  3. (1985). A point-source outbreak of campylobacteriosis associated with consumption of raw milk. doi
  4. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. doi
  5. (1999). An iron-regulated alkyl hydroperoxide reductase (AhpC) confers aerotolerance and oxidative stress resistance to the microaerophilic pathogen Campylobacter jejuni.
  6. (2003). Bacterial iron homeostasis. doi
  7. (1998). Binding of heme-hemopexin complexes by soluble HxuA protein allows utilization of this complexed heme by Haemophilus influenzae.
  8. (2001). Binding specificity of the Porphyromonas gingivalis heme and hemoglobin receptor HmuR, gingipain K, and gingipain R1 for heme, porphyrins, and metalloporphyrins. doi
  9. (2005). Campylobacter jejuni gene expression in response to iron limitation and the role of Fur.
  10. (2004). Campylobacter, from obscurity to celebrity. doi
  11. (2005). Carbohydrate mimicry: a new paradigm of autoimmune diseases. doi
  12. (2000). Characterization and expression of HmuR, a TonB-dependent hemoglobin receptor of Porphyromonas gingivalis. doi
  13. (2004). Characterization of heme uptake cluster genes in the fish pathogen Vibrio anguillarum. doi
  14. (1997). Characterization of hemin binding activity of Streptococcus pneumoniae.
  15. (1995). Characterization of the hgbA locus encoding a hemoglobin receptor from Haemophilus ducreyi.
  16. Characterization of the Plesiomonas shigelloides genes encoding the heme iron utilization system. doi
  17. (1994). Characterization of the Vibrio cholerae outer membrane heme transport protein HutA: sequence of the gene, regulation of expression, and homology to the family of TonB-dependent proteins.
  18. (1998). Cloning and characterization of tdhA, a locus encoding a TonB-dependent heme receptor from Haemophilus ducreyi.
  19. (1994). CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. doi
  20. (2005). Cytolethal distending toxin, doi
  21. (2000). Degradation of heme in gramnegative bacteria: the product of the hemO gene of neisseriae is a heme oxygenase. doi
  22. (1992). Differentiated caco-2 cells as a model for enteric invasion by Campylobacter jejuni and Campylobacter coli. doi
  23. (1999). Effect of multiple mutations in the hemoglobin- and hemoglobin-haptoglobin-binding proteins, HgpA, HgpB, and HgpC, of Haemophilus influenzae type b. doi
  24. (2001). Emerging strategies in microbial haem capture. doi
  25. (1998). Expression and characterization of a heme oxygenase (HmuO) from Corynebacterium diphtheriae. Iron acquisition requires oxidative cleavage of the heme macrocycle. doi
  26. (2004). Free and hemophorebound heme acquisitions through the outer membrane receptor HasR have different requirements for the TonB-ExbB-ExbD complex. doi
  27. (1994). Fur (ferric uptake regulation) protein interaction with target DNA: comparison of gel retardation, footprinting and electron microscopy analyses. doi
  28. (1998). Genetic manipulation of enteric Campylobacter species, doi
  29. (2005). Genetic variability of the heme uptake system among different strains of the fish pathogen Vibrio anguillarum: identification of a new heme receptor. doi
  30. (2000). Genetics and regulation of two distinct haem-uptake systems, phu and has, in Pseudomonas aeruginosa. doi
  31. (2001). Haem utilization in Vibrio cholerae involves multiple TonB-dependent haem receptors. doi
  32. (1993). Heat-labile and heat-stable haemolysins of Campylobacter jejuni. doi
  33. (1999). Heme degradation as catalyzed by a recombinant bacterial heme oxygenase (HmuO) from Corynebacterium diphtheriae. doi
  34. (2005). Heme oxygenase and heme degradation. doi
  35. (1992). Hemin uptake system of Yersinia enterocolitica: similarities with other TonB-dependent systems in gram-negative bacteria.
  36. (2004). HutZ is required for efficient heme utilization in Vibrio cholerae. doi
  37. (1995). Identification and purification of a conserved heme-regulated hemoglobin-binding outer membrane protein from Haemophilus ducreyi.
  38. (2004). Identification of a haem-utilization protein (Hup) in Haemophilus influenzae. doi
  39. (2005). Identification of an Escherichia coli O157:H7 heme oxygenase with tandem functional repeats. doi
  40. (2004). Identification of Campylobacter jejuni genes involved in commensal colonization of the chick gastrointestinal tract. doi
  41. (1998). Identification of Campylobacter jejuni promoter sequences.
  42. (2001). Involvement of HxuC outer membrane protein in utilization of hemoglobin by Haemophilus influenzae. doi
  43. (1992). Iron acquisition and hemolysin production by Campylobacter jejuni.
  44. (2004). Iron acquisition and regulation in Campylobacter jejuni. doi
  45. (2001). Iron and metal regulation in bacteria. doi
  46. (2004). Iron and microbial infection. doi
  47. (1998). Iron-responsive gene regulation in a Campylobacter jejuni Fur mutant.
  48. (1994). Iron-responsive genetic regulation in Campylobacter jejuni: cloning and characterization of a fur homolog.
  49. (2004). IsdG and IsdI, hemedegrading enzymes in the cytoplasm of Staphylococcus aureus. doi
  50. (1995). Molecular characterization of katA from Campylobacter jejuni and generation of a catalase-deficient mutant of Campylobacter coli by interspecific allelic exchange. doi
  51. (1999). Molecular characterization of the hemin uptake locus (hmu) from Yersinia pestis and analysis of hmu mutants for hemin and hemoprotein utilization.
  52. (2004). Molecular cloning of haemoglobin-binding protein HgbA in the outer membrane of Actinobacillus pleuropneumoniae. doi
  53. (2001). Molecular cloning: a laboratory manual, 3rd ed. doi
  54. (1994). Observation of binding and polymerization of Fur repressor onto operatorcontaining DNA with electron and atomic force microscopes. doi
  55. (1997). Pathogenesis of enteric infection by Campylobacter. doi
  56. (1978). Purification and properties of heme oxygenase from pig spleen microsomes.
  57. (2000). Reaction intermediates and single turnover rate constants for the oxidation of heme by human heme oxygenase-1. doi
  58. (1997). Signal transduction and transcriptional and posttranscriptional control of iron-regulated genes in bacteria.
  59. (1998). Structure of the Shigella dysenteriae haem transport locus and its phylogenetic distribution in enteric bacteria. doi
  60. (1983). Studies on transformation of Escherichia coli with plasmids. doi
  61. The genome sequence of the food-borne pathogen Campylobacter jejuni reveals hypervariable sequences.
  62. (2001). The iron-induced ferredoxin FdxA of Campylobacter jejuni is involved in aerotolerance. doi
  63. (2000). The iron-responsive regulator Fur of Campylobacter jejuni is expressed from two separate promoters. doi
  64. (1995). The Neisseria meningitidis haemoglobin receptor: its role in iron utilization and virulence. doi
  65. (2002). The role of iron in Campylobacter gene regulation, metabolism and oxidative stress defense. doi
  66. (1994). Transport of haemin across the cytoplasmic membrane through a haemin-specific periplasmic binding-proteindependent transport system in Yersinia enterocolitica. doi
  67. (1998). Transport of intact porphyrin by HpuAB, the hemoglobin-haptoglobin utilization system of Neisseria meningitidis.
  68. (1986). Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. doi
  69. (2005). Use of genome-wide expression profiling and mutagenesis to study the intestinal lifestyle of Campylobacter jejuni. doi
  70. (2000). Use of heme compounds as iron sources by pathogenic neisseriae requires the product of the hemO gene. doi

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