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Infection and Immunity, May 2000, p. 2594-2601, Vol. 68, No. 5
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

The galE Gene of Campylobacter jejuni Is Involved in Lipopolysaccharide Synthesis and Virulence

Benjamin N. Fry,1,* Shi Feng,1 Yuen-Yuen Chen,1 Diane G. Newell,2 Peter J. Coloe,1 and Victoria Korolik1,dagger

Department of Applied Biology and Biotechnology, Royal Melbourne Institute of Technology University, Melbourne 3001, Victoria Australia,1 and Veterinary Laboratories Agency, Weybridge, United Kingdom2

Received 1 November 1999/Returned for modification 21 December 1999/Accepted 27 January 2000

Lipopolysaccharide (LPS) is one of the main virulence factors of gram-negative bacteria. The LPS from Campylobacter spp. has endotoxic properties and has been shown to play a role in adhesion. We previously cloned a gene cluster (wla) which is involved in the synthesis of the Campylobacter jejuni 81116 LPS molecule. Sequence alignment of the first gene in this cluster indicated similarity with galE genes. These genes encode a UDP-glucose 4-epimerase, which catalyzes the interconversion of UDP-galactose and UDP-glucose. A Salmonella galE mutant was transformed with the galE gene from C. jejuni. The LPS analysis of wild-type, galE, and complemented galE Salmonella strains showed that the C. jejuni galE gene could restore the smooth wild-type Salmonella LPS. A UDP-glucose 4-epimerase assay was used to demonstrate that the galE gene from C. jejuni encoded this epimerase. We constructed a C. jejuni galE mutant which expressed a lipid A-core molecule of reduced molecular weight that did not react with antiserum raised against the parental strain. These results show an essential role for the galE gene in the synthesis of C. jejuni LPS. The galE mutant also showed a reduction in its ability to adhere to and invade INT407 cells. However, it was still able to colonize chickens to the same level as the wild-type strain. The serum resistance and hemolytic activity of this mutant were not changed compared to the parent strain. The ability of the mutant to take up DNA and integrate it in its genome was reduced 20-fold. These results show that LPS of C. jejuni is an important virulence factor.

* Corresponding author. Mailing address: Department of Applied Biology and Biotechnology, RMIT-University, GPO Box 2476V, Melbourne 3001, Victoria, Australia. Phone: 61 3 99253366. Fax: 61 3 96623421. E-mail: ben.fry{at}rmit.edu.au.

dagger Present address: School of Health Science, Griffith University, Gold Coast, QLD 4217, Australia.

Infection and Immunity, May 2000, p. 2594-2601, Vol. 68, No. 5
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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