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Journal of Bacteriology, November 2009, p. 6950-6959, Vol. 191, No. 22
0021-9193/09/$08.00+0     doi:10.1128/JB.00787-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Structural and Genetic Basis for the Serological Differentiation of Pasteurella multocida Heddleston Serotypes 2 and 5 {triangledown}

Frank St. Michael,1,{dagger} Marina Harper,2,{dagger} Henrietta Parnas,1 Marietta John,2 Jacek Stupak,1 Evgeny Vinogradov,1 Ben Adler,2 John D. Boyce,2 and Andrew D. Cox1*

Institute for Biological Sciences, National Research Council, Ottawa, Ontario, Canada K1A 0R6,1 Australian Research Council Centre of Excellence in Structural and Functional Microbial Genomics, Department of Microbiology, Monash University, Victoria 3800, Australia2

Received 16 June 2009/ Accepted 14 September 2009

Pasteurella multocida is classified into 16 serotypes according to the Heddleston typing scheme. As part of a comprehensive study to define the structural and genetic basis of this scheme, we have determined the structure of the lipopolysaccharide (LPS) produced by P. multocida strains M1404 (B:2) and P1702 (E:5), the type strains for serotypes 2 and 5, respectively. The only difference between the LPS structures made by these two strains was the absence of a phosphoethanolamine (PEtn) moiety at the 3 position of the second heptose (Hep II) in M1404. Analysis of the lpt-3 gene, required for the addition of this PEtn residue, revealed that the gene was intact in P1702 but contained a nonsense mutation in M1404. Expression of an intact copy of lpt-3 in M1404 resulted in the attachment of a PEtn residue to the 3 position of the Hep II residue, generating an LPS structure identical to that produced by P1702. We identified and characterized each of the glycosyltransferase genes required for assembly of the serotype 2 and 5 LPS outer core. Monoclonal antibodies raised against serotype 2 LPS recognized the serotype 2/5-specific outer core LPS structure, but recognition of this structure was inhibited by the PEtn residue on Hep II. These data indicate that the serological classification of strains into Heddleston serotypes 2 and 5 is dependent on the presence or absence of PEtn on Hep II.

* Corresponding author. Mailing address: Institute for Biological Sciences, 100 Sussex Drive, National Research Council, Ottawa, Ontario K1A 0R6, Canada. Phone: (613) 991-6172. Fax: (613) 952-9092. E-mail: Andrew.Cox{at}nrc-cnrc.gc.ca

{triangledown} Published ahead of print on 18 September 2009.

{dagger} F.S.M. and M.H. contributed equally to this work.

Journal of Bacteriology, November 2009, p. 6950-6959, Vol. 191, No. 22
0021-9193/09/$08.00+0     doi:10.1128/JB.00787-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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