Bacteriophage F336 Recognizes the Capsular Phosphoramidate Modification of Campylobacter jejuni NCTC11168

Author:

Sørensen Martine C. Holst1,van Alphen Lieke B.2,Harboe Anne1,Li Jianjun3,Christensen Bjarke Bak4,Szymanski Christine M.2,Brøndsted Lone1

Affiliation:

1. Department of Veterinary Disease Biology, University of Copenhagen, Stigbøjlen 4, 1870 Frederiksberg, Denmark

2. Department of Biological Sciences, Alberta Innovates Centre for Carbohydrate Science, University of Alberta, 11455 Saskatchewan Drive, Edmonton, Alberta T6G 2E9, Canada

3. Institute for Biological Sciences, National Research Council of Canada, 100 Sussex Drive, Ottawa, Ontario K1A OR6, Canada

4. National Food Institute, Technical University of Denmark, Mørkhøj Bygade 19, 2860 Søborg, Denmark

Abstract

ABSTRACT Bacteriophages infecting the food-borne human pathogen Campylobacter jejuni could potentially be exploited to reduce bacterial counts in poultry prior to slaughter. This bacterium colonizes the intestinal tract of poultry in high numbers, and contaminated poultry meat is regarded as the major source of human campylobacteriosis. In this study, we used phage F336 belonging to the Myoviridae family to select a C. jejuni NCTC11168 phage-resistant strain, called 11168R, with the aim of investigating the mechanisms of phage resistance. We found that phage F336 has reduced adsorption to 11168R, thus indicating that the receptor is altered. While proteinase K-treated C. jejuni cells did not affect adsorption, periodate treatment resulted in reduced adsorption, suggesting that the phage binds to a carbohydrate moiety. Using high-resolution magic angle spinning nuclear magnetic resonance (NMR) spectroscopy, we found that 11168R lacks an O -methyl phosphoramidate (MeO P N) moiety attached to the Gal f NAc on the capsular polysaccharide (CPS), which was further confirmed by mass spectroscopy. Sequence analysis of 11168R showed that the potentially hypervariable gene cj1421 , which encodes the Gal f NAc MeO P N transferase, contains a tract of 10 Gs, resulting in a nonfunctional gene product. However, when 11168R reverted back to phage sensitive, cj1421 contained 9 Gs, and the Gal f NAc MeO P N was regained in this strain. In summary, we have identified the phase-variable MeO P N moiety, a common component of the diverse capsular polysaccharides of C. jejuni , as a novel receptor of phages infecting this bacterium.

Publisher

American Society for Microbiology

Subject

Molecular Biology,Microbiology

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