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Infection and Immunity, October 2001, p. 6225-6230, Vol. 69, No. 10
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.10.6225-6230.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Ferrochelatase Is Present in Brucella abortus and Is Critical for Its Intracellular Survival and Virulence

Marta Almirón,1 Marcela Martínez,1 Norberto Sanjuan,2 and Rodolfo A. Ugalde1,*

Instituto de Investigaciones Biotecnológicas, Instituto Tecnológico de Chascomús (IIB, INTECH), Consejo Nacional de Investigaciones Científicas y Técnicas, Universidad Nacional de General San Martín (CONICET-UNSAM),1 and Laboratorio de Patología Experimental, Departamento de Microbiología, Facultad de Medicina, Universidad de Buenos Aires,2 Buenos Aires, Argentina

Received 23 February 2001/Returned for modification 9 May 2001/Accepted 25 June 2001

Brucella spp. are pathogenic bacteria that cause brucellosis, an animal disease which can also affect humans. Although understanding the pathogenesis is important for the health of animals and humans, little is known about virulence factors associated with it. In order for chronic disease to be established, Brucella spp. have developed the ability to survive inside phagocytes by evading cell defenses. It hides inside vacuoles, where it then replicates, indicating that it has an active metabolism. The purpose of this work was to obtain better insight into the intracellular metabolism of Brucella abortus. During a B. abortus genomic sequencing project, a clone coding a putative gene homologous to hemH was identified and sequenced. The amino acid sequence revealed high homology to members of the ferrochelatase family. A knockout mutant displayed auxotrophy for hemin, defective intracellular survival inside J774 and HeLa cells, and lack of virulence in BALB/c mice. This phenotype was overcome by complementing the mutant strain with a plasmid harboring wild-type hemH. These data demonstrate that B. abortus synthesizes its own heme and also has the ability to use an external source of heme; however, inside cells, there is not enough available heme to support its intracellular metabolism. It is concluded that ferrochelatase is essential for the multiplication and intracellular survival of B. abortus and thus for the establishment of chronic disease as well.

* Corresponding author. Mailing address: Instituto de Investigaciones Biotecnológicas, Av. General Paz entre Constituyentes y Albarellos, 1650 Buenos Aires, Argentina. Phone: (5411) 4580 7255. Fax: (5411) 4752 9639. E-mail: rugalde{at}inti.gov.ar.

Infection and Immunity, October 2001, p. 6225-6230, Vol. 69, No. 10
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.10.6225-6230.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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