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Infection and Immunity, December 2003, p. 6892-6898, Vol. 71, No. 12
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.12.6892-6898.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Identification and Characterization of pptA: a Gene Involved in the Phase-Variable Expression of Phosphorylcholine on Pili of Neisseria meningitidis

Matthew J. Warren and Michael P. Jennings*

Department of Microbiology and Parasitology, School of Molecular and Microbial Sciences, University of Queensland, Brisbane, Queensland 4072, Australia

Received 21 May 2003/ Returned for modification 26 June 2003/ Accepted 3 September 2003

Pili of pathogenic Neisseria are major virulence factors associated with adhesion, cytotoxicity, twitching motility, autoaggregation, and DNA transformation. Pili are modified posttranslationally by the addition of phosphorylcholine. However, no genes involved in either the biosynthesis or the transfer of phosphorylcholine in Neisseria meningitidis have been identified. In this study, we identified five candidate open reading frames (ORFs) potentially involved in the biosynthesis or transfer of phosphorylcholine to pilin in N. meningitidis. Insertional mutants were constructed for each ORF in N. meningitidis strain C311#3 to determine their effect on phosphorylcholine expression. The effect of the mutant ORFs on the modification by phosphorylcholine was analyzed by Western analysis with phosphorylcholine-specific monoclonal antibody TEPC-15. Analysis of the mutants showed that ORF NMB0415, now defined as pptA (pilin phosphorylcholine transferase A), is involved in the addition of phosphorylcholine to pilin in N. meningitidis. Additionally, the phase variation (high frequency on-off switching of expression) of phosphorylcholine on pilin is due to changes in a homopolymeric guanosine tract in pptA.

* Corresponding author. Mailing address: Department of Microbiology and Parasitology, School of Molecular and Microbial Sciences, University of Queensland, Brisbane, Queensland 4072, Australia. Phone: 61 7 3365 4879. Fax: 61 7 3365 4620. E-mail: jennings{at}uq.edu.au.

Editor: J. N. Weiser

Infection and Immunity, December 2003, p. 6892-6898, Vol. 71, No. 12
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.12.6892-6898.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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