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Journal of Bacteriology, August 2005, p. 5156-5165, Vol. 187, No. 15
0021-9193/05/$08.00+0     doi:10.1128/JB.187.15.5156-5165.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Identification of a D-glycero-D-manno-Heptosyltransferase Gene from Helicobacter pylori

Koji Hiratsuka, Susan M. Logan, J. Wayne Conlan, Vandana Chandan, Annie Aubry, Natalia Smirnova, Heather Ulrichsen, Kenneth H. N. Chan, Douglas W. Griffith, Blair A. Harrison, Jianjun Li, and Eleonora Altman*

Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario, Canada K1A 0R6

Received 25 January 2005/ Accepted 27 April 2005

We have identified a Helicobacter pylori D-glycero-D-manno-heptosyltransferase gene, HP0479, which is involved in the biosynthesis of the outer core region of H. pylori lipopolysaccharide (LPS). Insertional inactivation of HP0479 resulted in formation of a truncated LPS molecule lacking an {alpha}-1,6-glucan-, DD-heptose-containing outer core region and O-chain polysaccharide. Detailed structural analysis of purified LPS from HP0479 mutants of strains SS1, 26695, O:3, and PJ1 by a combination of chemical and mass spectrometric methods showed that HP0479 likely encodes {alpha}-1,2-D-glycero-D-manno-heptosyltransferase, which adds a D-glycero-D-manno-heptose residue (DDHepII) to a distal DD-heptose of the core oligosaccharide backbone of H. pylori LPS. When the wild-type HP0479 gene was reintegrated into the chromosome of strain 26695 by using an "antibiotic cassette swapping" method, the complete LPS structure was restored. Introduction of the HP0479 mutation into the H. pylori mouse-colonizing Sydney (SS1) strain and the clinical isolate PJ1, which expresses DD-heptoglycan, resulted in the loss of colonization in a mouse model. This indicates that H. pylori expressing a deeply truncated LPS is unable to successfully colonize the murine stomach and provides evidence for a critical role of the outer core region of H. pylori LPS in colonization.

* Corresponding author. Mailing address: Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario, Canada K1A 0R6. Phone: (613) 990-0904. Fax: (613) 941-1327. E-mail: Eleonora.altman{at}nrc-cnrc.gc.ca.

Journal of Bacteriology, August 2005, p. 5156-5165, Vol. 187, No. 15
0021-9193/05/$08.00+0     doi:10.1128/JB.187.15.5156-5165.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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