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Infection and Immunity, January 2006, p. 108-117, Vol. 74, No. 1
0019-9567/06/$08.00+0     doi:10.1128/IAI.74.1.108-117.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

The Brucella suis Type IV Secretion System Assembles in the Cell Envelope of the Heterologous Host Agrobacterium tumefaciens and Increases IncQ Plasmid pLS1 Recipient Competence

Anna Carle,1 Christoph Höppner,1 Khaled Ahmed Aly,2 Qing Yuan,2 Amke den Dulk-Ras,3 Annette Vergunst,3,4 David O'Callaghan,4 and Christian Baron1,2*

Department Biologie I, Bereich Mikrobiologie, Ludwig-Maximilians-Universität, Maria-Ward-Str. 1a, D-80638 Munich, Germany,1 Department of Biology, McMaster University, 1280 Main St. West, Hamilton, Ontario LS8 4K1, Canada,2 Clusius Laboratory, Institute of Biology, Leiden University, Wassenaarseweg 64, 2333 AL Leiden, The Netherlands,3 Unité 431, UFR de Médecine, Institut National de la Santé et de la Recherche Médicale, CS 83021, Avenue J. F. Kennedy, 30908 Nîmes Cedex, France4

Received 10 August 2005/ Returned for modification 22 September 2005/ Accepted 23 September 2005

Pathogenic Brucella species replicate within mammalian cells, and their type IV secretion system is essential for intracellular survival and replication. The options for biochemical studies on the Brucella secretion system are limited due to the rigidity of the cells and biosafety concerns, which preclude large-scale cell culture and fractionation. To overcome these problems, we heterologously expressed the Brucella suis virB operon in the closely related {alpha}2-proteobacterium Agrobacterium tumefaciens and showed that the VirB proteins assembled into a complex. Eight of the twelve VirB proteins were detected in the membranes of the heterologous host with specific antisera. Cross-linking indicated protein-protein interactions similar to those in other type IV secretion systems, and the results of immunofluorescence analysis supported the formation of VirB protein complexes in the cell envelope. Production of a subset of the B. suis VirB proteins (VirB3-VirB12) in A. tumefaciens strongly increased its ability to receive IncQ plasmid pLS1 in conjugation experiments, and production of VirB1 further enhanced the conjugation efficiency. Plasmid recipient competence correlated with periplasmic leakage and the detergent sensitivity of A. tumefaciens, suggesting a weakening of the cell envelope. Heterologous expression thus permits biochemical characterization of B. suis type IV secretion system assembly.

* Corresponding author. Mailing address: McMaster University, Department of Biology, 1280 Main St. West, Hamilton, ON LS8 4K1, Canada. Phone: (905) 525-9140, x26692. Fax: (905) 522-6066. E-mail: baronc{at}mcmaster.ca.

Editor: D. L. Burns

Infection and Immunity, January 2006, p. 108-117, Vol. 74, No. 1
0019-9567/06/$08.00+0     doi:10.1128/IAI.74.1.108-117.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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