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Infection and Immunity, June 2004, p. 3436-3443, Vol. 72, No. 6
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.6.3436-3443.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

A Heptosyltransferase Mutant of Pasteurella multocida Produces a Truncated Lipopolysaccharide Structure and Is Attenuated in Virulence

Marina Harper,1,2 Andrew D. Cox,3 Frank St. Michael,3 Ian W. Wilkie,4 John D. Boyce,1,2 and Ben Adler1,2*

Australian Bacterial Pathogenesis Program, Department of Microbiology,1 Australian Research Council Centre for Structural and Functional Microbial Genomics, Monash University, Melbourne, Victoria 3800,2 Veterinary Pathology and Anatomy, University of Queensland, Brisbane, Queensland 4072, Australia,4 Institute for Biological Sciences, National Research Council, Ottawa, Canada3

Received 11 November 2003/ Returned for modification 10 February 2004/ Accepted 27 February 2004

Pasteurella multocida is the causative agent of fowl cholera in birds. In a previous study using signature-tagged mutagenesis, we identified a mutant, AL251, which was attenuated for virulence in mice and in the natural chicken host. Sequence analysis indicated that AL251 had an insertional inactivation of the gene waaQPM, encoding a putative heptosyl transferase, required for the addition of heptose to lipopolysaccharide (LPS) (M. Harper, J. D. Boyce, I. W. Wilkie, and B. Adler, Infect. Immun. 71:5440-5446, 2003). In the present study, using mass spectrometry and nuclear magnetic resonance, we have confirmed the identity of the enzyme encoded by waaQPM as a heptosyl transferase III and demonstrated that the predominant LPS glycoforms isolated from this mutant are severely truncated. Complementation experiments demonstrated that providing a functional waaQPM gene in trans can restore both the LPS to its full length and growth in mice to wild-type levels. Furthermore, we have shown that mutant AL251 is unable to cause fowl cholera in chickens and that the attenuation observed is not due to increased serum sensitivity.

* Corresponding author. Mailing address: Department of Microbiology, Monash University, Clayton, Victoria 3800, Australia. Phone: 61 03 9905 4815. Fax: 61 03 9905 4811. E-mail: ben.adler{at}med.monash.edu.au.

Editor: V. J. DiRita

Infection and Immunity, June 2004, p. 3436-3443, Vol. 72, No. 6
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.6.3436-3443.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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