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Journal of Bacteriology, May 2008, p. 3129-3139, Vol. 190, No. 9
0021-9193/08/$08.00+0     doi:10.1128/JB.01830-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Expression of the Gonococcal Global Regulatory Protein Fur and Genes Encompassing the Fur and Iron Regulon during In Vitro and In Vivo Infection in Women{triangledown} ,{dagger}

Sarika Agarwal,1,2 Shite Sebastian,1,{ddagger} Borys Szmigielski,1,§ Peter A. Rice,2 and Caroline A. Genco1,3*

Department of Medicine, Section of Molecular Medicine,1 Department of Microbiology, Boston University School of Medicine, Boston, Massachusetts 02118,3 Department of Medicine, Division of Infectious Diseases and Immunology, University of Massachusetts Medical School, Worcester, Massachusetts 016052

Received 20 November 2007/ Accepted 17 February 2008

The ferric uptake regulatory protein, Fur, functions as a global regulatory protein of gene transcription in the mucosal pathogen Neisseria gonorrhoeae. We have shown previously that several N. gonorrhoeae Fur-repressed genes are expressed in vivo during mucosal gonococcal infection in men, which suggests that this organism infects in an iron-limited environment and that Fur is expressed under these conditions. In this study we have demonstrated expression of the gonococcal fur gene in vitro, in human cervical epithelial cells, and in specimens from female subjects with uncomplicated gonococcal infection. In vitro studies confirmed that the expression of the gonococcal fur gene was repressed during growth under iron-replete growth conditions but that a basal level of the protein was maintained. Using GFP transcriptional fusions constructed from specific Fur binding sequences within the fur promoter/operator region, we determined that this operator region was functional during N. gonorrhoeae infection of cervical epithelial cells. Furthermore, reverse transcription-PCR analysis, as well as microarray analysis, using a custom Neisseria Fur and iron regulon microarray revealed that several Fur- and iron-regulated genes were expressed during N. gonorrhoeae infection of cervical epithelial cells. Microarray analysis of specimens obtained from female subjects with uncomplicated gonococcal infection corroborated our in vitro findings and point toward a key role of gonococcal Fur- and iron-regulated genes in gonococcal disease.

* Corresponding author. Mailing address: Department of Medicine, Section of Molecular Medicine, 650 Albany St., Rm. 314, Boston University School of Medicine, Boston, MA 02118. Phone: (617) 414-5305. Fax: (617) 414-1719. E-mail: caroline.genco{at}bmc.org

{triangledown} Published ahead of print on 29 February 2008.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

{ddagger} Present address: SanofiPasteur, Discovery Drive, Swiftwater, PA 18370.

§ Present address: Department of Microbiology, Faculty of Biotechnology, Jagiellonian University, Gronostajowa 8, Kraków, Poland.

Journal of Bacteriology, May 2008, p. 3129-3139, Vol. 190, No. 9
0021-9193/08/$08.00+0     doi:10.1128/JB.01830-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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