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J. Biol. Chem., Vol. 279, Issue 19, 20327-20338, May 7, 2004

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Genome-wide Expression Analyses of Campylobacter jejuni NCTC11168 Reveals Coordinate Regulation of Motility and Virulence by flhA^*

Catherine D. Carrillo, Eduardo Taboada, John H. E. Nash, Patricia Lanthier, John Kelly, Peter C. Lau, Rachel Verhulp, Oksana Mykytczuk, Jonathan Sy, Wendy A. Findlay, Kingsley Amoako, Susantha Gomis¶, Philip Willson, John W. Austin||, Andy Potter, Lorne Babiuk, Brenda Allan**, and Christine M. Szymanski

From the Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario K1A 0R6, Canada, Vaccine and Infectious Disease Organization, Saskatoon, Saskatchewan S7N 5E3, Canada, the ^¶Department of Veterinary Pathology, Western College of Medicine, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5B4, Canada, and the ^||Bureau of Microbial Hazards, Health Canada, Ottawa, Ontario K1A 0L2, Canada

We examined two variants of the genome-sequenced strain, Campylobacter jejuni NCTC11168, which show marked differences in their virulence properties including colonization of poultry, invasion of Caco-2 cells, and motility. Transcript profiles obtained from whole genome DNA microarrays and proteome analyses demonstrated that these differences are reflected in late flagellar structural components and in virulence factors including those involved in flagellar glycosylation and cytolethal distending toxin production. We identified putative ²⁸ and ⁵⁴ promoters for many of the affected genes and found that greater differences in expression were observed for ²⁸-controlled genes. Inactivation of the gene encoding ²⁸, fliA, resulted in an unexpected increase in transcripts with ⁵⁴ promoters, as well as decreased transcription of ²⁸-regulated genes. This was unlike the transcription profile observed for the attenuated C. jejuni variant, suggesting that the reduced virulence of this organism was not entirely due to impaired function of ²⁸. However, inactivation of flhA, an important component of the flagellar export apparatus, resulted in expression patterns similar to that of the attenuated variant. These findings indicate that the flagellar regulatory system plays an important role in campylobacter pathogenesis and that flhA is a key element involved in the coordinate regulation of late flagellar genes and of virulence factors in C. jejuni.

Received for publication, February 2, 2004

^* This work was supported by the National Research Council Genomics and Health Initiative (to C. D. C., E. T., J. H. E. N., P. L., J. K., P. C. L., R. V., O. M., J. S., W. A. F., and C. M. S.), by the Canadian Bacterial Diseases Network Centre of Excellence and Genome Canada/Genome Prairie (to A. P. and L. B.), and by the Poultry Industry Council, Agricultural Development Fund, Alberta Poultry Producers, and OMAF Food Safety Research Program (to B. A.). This work is published with the permission of the Director of the VIDO as journal series number 353. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains six additional tables.

^** To whom correspondence may be addressed: Vaccine and Infectious Disease Organization, 120 Veterinary Rd., Saskatoon, Saskatchewan S7N 5E3, Canada. Tel.: 306-966-7486; Fax: 306-966-7478; E-mail: allanb{at}sask.usask.ca.

To whom correspondence may be addressed: Institute for Biological Sciences, National Research Council of Canada, 100 Sussex Dr., Ottawa, Ontario K1A 0R6, Canada. Tel.: 613-991-4342; Fax: 613-952-9092; E-mail: christine.szymanski{at}nrc-cnrc.gc.ca.

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