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

Infection and Immunity - Tập 72 Số 6 - Trang 3436-3443 - 2004
Marina Harper1,2, Andrew D. Cox3, Frank St. Michael3, Ian Wilkie4, John D. Boyce1,2, Ben Adler1,2
1Australian Bacterial Pathogenesis Program, Department of Microbiology
2Australian Research Council Centre for Structural and Functional Microbial Genomics, Monash University, Melbourne, Victoria 3800
3Institute for Biological Sciences, National Research Council, Ottawa, Canada
4Veterinary Pathology and Anatomy, University of Queensland, Brisbane, Queensland 4072, Australia

Tóm tắt

ABSTRACT 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 waaQ PM , 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 waaQ PM 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 waaQ PM 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.

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Infection and Immunity

Tập 72 Số 6

3436-3443

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