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Journal of Bacteriology, November 2006, p. 7862-7875, Vol. 188, No. 22
0021-9193/06/$08.00+0     doi:10.1128/JB.00994-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Heme Utilization in Campylobacter jejuni{triangledown}

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

Department of Genetics, Adrian Building, University Road, University of Leicester, Leicester LE1 7RH, United Kingdom

Received 7 July 2006/ Accepted 8 September 2006

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.

* Corresponding author. Mailing address: Department of Genetics, Adrian Building, University Road, University of Leicester, Leicester LE1 7RH, United Kingdom. Phone: (44) 1162523434. Fax: (44) 1162523378. E-mail: ket{at}leicester.ac.uk.

{triangledown} Published ahead of print on 15 September 2006.

Journal of Bacteriology, November 2006, p. 7862-7875, Vol. 188, No. 22
0021-9193/06/$08.00+0     doi:10.1128/JB.00994-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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