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Journal of Bacteriology, October 2007, p. 7384-7391, Vol. 189, No. 20
0021-9193/07/$08.00+0     doi:10.1128/JB.00948-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Decoration of Pasteurella multocida Lipopolysaccharide with Phosphocholine Is Important for Virulence{triangledown}

Marina Harper,1 Andrew Cox,2 Frank St. Michael,2 Henrietta Parnas,2 Ian Wilkie,3 P. J. Blackall,4 Ben Adler,1* and John D. Boyce1

Australian Research Council Centre of Excellence in Structural and Functional Microbial Genomics, Department of Microbiology, Monash University, Victoria 3800, Australia,1 Institute for Biological Sciences, National Research Council, Ottawa, Canada K1A OR6,2 Veterinary Pathology and Anatomy, University of Queensland, Brisbane, Queensland 4072, Australia,3 Department of Primary Industries and Fisheries (Queensland) Animal Research Institute, Yeerongpilly, Queensland 4105, Australia4

Received 15 June 2007/ Accepted 9 August 2007

Phosphocholine (PCho) is an important substituent of surface structures expressed by a number of bacterial pathogens. Its role in virulence has been investigated in several species, in which it has been shown to play a role in bacterial adhesion to mucosal surfaces, in resistance to antimicrobial peptides, or in sensitivity to complement-mediated killing. The lipopolysaccharide (LPS) structure of Pasteurella multocida strain Pm70, whose genome sequence is known, has recently been determined and does not contain PCho. However, LPS structures from the closely related, virulent P. multocida strains VP161 and X-73 were shown to contain PCho on their terminal galactose sugar residues. To determine if PCho was involved in the virulence of P. multocida, we used subtractive hybridization of the VP161 genome against the Pm70 genome to identify a four-gene locus (designated pcgDABC) which we show is required for the addition of the PCho residues to LPS. The proteins predicted to be encoded by pcgABC showed identity to proteins involved in choline uptake, phosphorylation, and nucleotide sugar activation of PCho. We constructed a P. multocida VP161 pcgC mutant and demonstrated that this strain produces LPS that lacks PCho on the terminal galactose residues. This pcgC mutant displayed reduced in vivo growth in a chicken infection model and was more sensitive to the chicken antimicrobial peptide fowlicidin-1 than the wild-type P. multocida strain.


* Corresponding author. Mailing address: Australian Research Council Centre of Excellence in Structural and Functional Microbial Genomics, Monash University, VIC 3800, Australia. Phone: 61 3 9905-4823. Fax: 61 3 9905-4811. E-mail: Ben.Adler{at}med.monash.edu.au

{triangledown} Published ahead of print on 17 August 2007.


Journal of Bacteriology, October 2007, p. 7384-7391, Vol. 189, No. 20
0021-9193/07/$08.00+0     doi:10.1128/JB.00948-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.







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