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Infection and Immunity, September 2005, p. 5554-5567, Vol. 73, No. 9
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.9.5554-5567.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Unusual Genetic Organization of a Functional Type I Protein Secretion System in Neisseria meningitidis

Karl G. Wooldridge,* Murat Kizil,{dagger} Damien B. Wells, and Dlawer A. A. Ala'Aldeen

Molecular Bacteriology and Immunology Group, Institute of Infection, Immunity and Inflammation, University of Nottingham, Nottingham NG7 2RD, United Kingdom

Received 8 November 2004/ Returned for modification 20 January 2005/ Accepted 19 April 2005

Proteins secreted by Neisseria meningitidis are thought to play important roles in the pathogenesis of meningococcal disease. These proteins include the iron-repressible repeat-in-toxin (RTX) exoprotein FrpC. Related proteins in other pathogens are secreted via a type I secretion system (TOSS), but such a system has not been demonstrated in N. meningitidis. An in silico search of the group B meningococcal genome suggested the presence of a uniquely organized TOSS. Genes encoding homologs of the Escherichia coli HlyB (ATP-binding), HlyD (membrane fusion), and TolC (outer membrane channel) proteins were identified. In contrast to the cistronic organization of the secretion genes in most other rtx operons, the hlyD and tolC genes were adjacent but unlinked to hlyB; neither locus was part of an operon containing genes encoding putative TOSS substrates. Both loci were flanked by genes normally associated with mobile genetic elements. The three genes were shown to be expressed independently. Mutation at either locus resulted in an inability to secrete FrpC and a related protein, here called FrpC2. Successful complementation of these mutations at an ectopic site confirmed the observed phenotypes were caused by loss of function of the putative TOSS genes. We show that genes scattered in the meningococcal genome encode a functional TOSS required for secretion of the meningococcal RTX proteins.

* Corresponding author. Mailing address: Division of Microbiology and Infectious Diseases, Queens Medical Centre, Nottingham NG7 2UH, United Kingdom. Phone: 44-115-9709162. Fax: 44-115-9709233. E-mail: karl.wooldridge{at}nottingham.ac.uk.

Editor: V. J. DiRita

{dagger} Present address: Department of Chemistry, Faculty of Science and Art, University of Dicle, 21280 Diyarbakir, Turkey.

Infection and Immunity, September 2005, p. 5554-5567, Vol. 73, No. 9
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.9.5554-5567.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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