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Journal of Bacteriology, September 2000, p. 4849-4855, Vol. 182, No. 17
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

A Homologue of an Operon Required for DNA Transfer in Agrobacterium Is Required in Brucella abortus for Virulence and Intracellular Multiplication

Rodrigo Sieira,1 Diego J. Comerci,1,2 Daniel O. Sánchez,1 and Rodolfo A. Ugalde1,*

Instituto de Investigaciones Biotecnológicas, Universidad Nacional de General San Martín, CONICET, San Martín 1650,1 and Comisión Nacional de Energía Atómica, División Agropecuaria, Centro Atómico Ezeiza 1804,2 Buenos Aires, Argentina

Received 2 March 2000/Accepted 16 June 2000

As part of a Brucella abortus 2308 genome project carried out in our laboratory, we identified, cloned, and sequenced a genomic DNA fragment containing a locus (virB) highly homologous to bacterial type IV secretion systems. The B. abortus virB locus is a collinear arrangement of 13 open reading frames (ORFs). Between virB1 and virB2 and downstream of ORF12, two degenerated, palindromic repeat sequences characteristic of Brucella intergenic regions were found. Gene reporter studies demonstrated that the B. abortus virB locus constitutes an operon transcribed from virB1 which is turned on during the stationary phase of growth. A B. abortus polar virB1 mutant failed to replicate in HeLa cells, indicating that the virB operon plays a critical role in intracellular multiplication. Mutants with polar and nonpolar mutations introduced in virB10 showed different behaviors in mice and in the HeLa cell infection assay, suggesting that virB10 per se is necessary for the correct function of this type IV secretion apparatus. Mouse infection assays demonstrated that the virB operon constitutes a major determinant of B. abortus virulence. It is suggested that putative effector molecules secreted by this type IV secretion system determine routing of B. abortus to an endoplasmic reticulum-related replication compartment.

* Corresponding author. Mailing address: Instituto de Investigaciones Biotecnológicas, Avenida General Paz entre Constituyentes y Albarellos, 1650, Buenos Aires, Argentina. Phone: (54-11) 4580 7255. Fax: (54-11) 4752 9639. E-mail: rugalde{at}inti.gov.ar.

Journal of Bacteriology, September 2000, p. 4849-4855, Vol. 182, No. 17
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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