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Journal of Bacteriology, July 2008, p. 5020-5030, Vol. 190, No. 14
0021-9193/08/$08.00+0     doi:10.1128/JB.00377-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Genetic and Functional Characterization of the Type IV Secretion System in Wolbachia{triangledown} ,{dagger}

Edwige Rancès, Denis Voronin, Van Tran-Van, and Patrick Mavingui*

Université de Lyon, Lyon, F-69003, France; Université Lyon 1, Lyon, F-69003, France; CNRS, UMR 5557, Ecologie Microbienne, Villeurbanne, F-69622, France; and IFR 41, Villeurbanne, F-69622, France

Received 14 March 2008/ Accepted 13 May 2008

A type IV secretion system (T4SS) is used by many symbiotic and pathogenic intracellular bacteria for the successful infection of and survival, proliferation, and persistence within hosts. In this study, the presence and function of the T4SS in Wolbachia strains were investigated by a combination of genetic screening and immunofluorescence microscopy. Two operons of virB-virD4 loci were found in the genome of Wolbachia pipientis strain wAtab3, from the Hymenoptera Asobara tabida, and strain wRi, infecting Drosophila simulans. One operon consisted of five vir genes (virB8, virB9, virB10, virB11, and virD4) and the downstream wspB locus. The other operon was composed of three genes (virB3, virB4, and virB6) and included four additional open reading frames (orf1 to orf4) orientated in the same direction. In cell culture and insect hosts infected with different Wolbachia strains, the bona fide vir genes were polycistronically transcribed, together with the downstream adjacent loci, notably, as virB8 to virD4 and wspB and as virB3, virB4, virB6, and orf1 to orf4. Two peptides encompassing conserved C and N termini of the Wolbachia VirB6 protein were used for the production of polyclonal antibodies. Anti-VirB6 antibodies could detect the corresponding recombinant protein by chemifluorescence on Western blots of total proteins from Escherichia coli transformants and Wolbachia strains cultured in cell lines. Using immunofluorescence microscopy, we further demonstrated that the VirB6 protein was produced by Wolbachia strains in ovaries of insects harboring wAtab3 or wRi and cell lines infected with wAlbB or wMelPop. As VirB6 is known to associate with other VirB proteins to form a membrane-spanning structure, this finding suggests that a T4SS may function in Wolbachia.

* Corresponding author. Mailing address: Écologie Microbienne UMR CNRS 5557-USC INRA 1193, Université Claude Bernard Lyon 1, Bât. G. Mendel, 43 boulevard du 11 Novembre 1918, 69622 Villeurbanne, France. Phone: (33) 472431143. Fax: (33) 472431223. E-mail: patrick.mavingui{at}univ-lyon1.fr

{triangledown} Published ahead of print on 23 May 2008.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

Journal of Bacteriology, July 2008, p. 5020-5030, Vol. 190, No. 14
0021-9193/08/$08.00+0     doi:10.1128/JB.00377-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Zouache, K., Voronin, D., Tran-Van, V., Mavingui, P. (2009). Composition of Bacterial Communities Associated with Natural and Laboratory Populations of Asobara tabida Infected with Wolbachia. Appl. Environ. Microbiol. 75: 3755-3764 [Abstract] [Full Text]  


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