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Infection and Immunity, December 2003, p. 7053-7060, Vol. 71, No. 12
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.12.7053-7060.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Attenuated Signature-Tagged Mutagenesis Mutants of Brucella melitensis Identified during the Acute Phase of Infection in Mice

P. Lestrate, A. Dricot, R.-M. Delrue, C. Lambert, V. Martinelli, X. De Bolle, J.-J. Letesson, and A. Tibor*

Unité de Recherche en Biologie Moléculaire (URBM), Laboratoire d'Immunologie et de Microbiologie, University of Namur, 5000 Namur, Belgium

Received 12 May 2003/ Returned for modification 17 June 2003/ Accepted 14 August 2003

For this study, we screened 1,152 signature-tagged mutagenesis mutants of Brucella melitensis 16M in a mouse model of infection and found 36 of them to be attenuated in vivo. Molecular characterization of transposon insertion sites showed that for four mutants, the affected genes were only present in Rhizobiaceae. Another mutant contained a disruption in a gene homologous to mosA, which is involved in rhizopine biosynthesis in some strains of Rhizobium, suggesting that this sugar may be involved in Brucella pathogenicity. A mutant was disrupted in a gene homologous to fliF, a gene potentially coding for the MS ring, a basal component of the flagellar system. Surprisingly, a mutant was affected in the rpoA gene, coding for the essential {alpha}-subunit of the RNA polymerase. This disruption leaves a partially functional protein, impaired for the activation of virB transcription, as demonstrated by the absence of induction of the virB promoter in the Tn5::rpoA background. The results presented here highlight the fact that the ability of Brucella to induce pathogenesis shares similarities with the molecular mechanisms used by both Rhizobium and Agrobacterium to colonize their hosts.

* Corresponding author. Mailing address: Laboratoire d'Immunologie et de Microbiologie, Unité de Recherche en Biologie Moléculaire (URBM), University of Namur, rue de Bruxelles 61, B-5000 Namur, Belgium. Phone: 3281724403. Fax: 3281724297. E-mail: Anne.tibor{at}fundp.ac.be.

Editor: D. L. Burns

Infection and Immunity, December 2003, p. 7053-7060, Vol. 71, No. 12
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.12.7053-7060.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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