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Infection and Immunity, October 2002, p. 5816-5821, Vol. 70, No. 10
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.10.5816-5821.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Coxiella burnetii Localizes in a Rab7-Labeled Compartment with Autophagic Characteristics

Walter Berón,1* Maximiliano G. Gutierrez,1 Michel Rabinovitch,2 and Maria I. Colombo1*

Instituto de Histología y Embriología, Facultad de Ciencias Médicas, Universidad Nacional de Cuyo-CONICET, Mendoza 5500, Argentina,1 Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, UNIFESP Sao Paulo, Sao Paulo, Brazil2

Received 29 March 2002/ Returned for modification 17 May 2002/ Accepted 17 June 2002

The obligate intracellular bacterium Coxiella burnetii, the agent of Q fever in humans and of coxiellosis in other animals, survives and replicates within large, acidified, phagolysosome-like vacuoles known to fuse homo- and heterotypically with other vesicles. To further characterize these vacuoles, HeLa cells were infected with C. burnetii phase II; 48 h later, bacteria-containing vacuoles were labeled by LysoTracker, a marker of acidic compartments, and accumulated monodansylcadaverine and displayed protein LC3, both markers of autophagic vacuoles. Furthermore, 3-methyladenine and wortmannin, agents known to inhibit early stages in the autophagic process, each blocked Coxiella vacuole formation. These autophagosomal features suggest that Coxiella vacuoles interact with the autophagic pathway. The localization and role of wild-type and mutated Rab5 and Rab7, markers of early and late endosomes, respectively, were also examined to determine the role of these small GTPases in the trafficking of C. burnetii phase II. Green fluorescent protein (GFP)-Rab5 and GFP-Rab7 constructs were overexpressed and visualized by fluorescence microscopy. Coxiella-containing large vacuoles were labeled with wild-type Rab7 (Rab7wt) and with GTPase-deficient mutant Rab7Q67L, whereas no colocalization was observed with the dominant-negative mutant Rab7T22N. The vacuoles were also decorated by GFP-Rab5Q79L but not by GFP-Rab5wt. These results suggest that Rab7 participates in the biogenesis of the parasitophorous vacuoles.

* Corresponding author. Mailing address: Instituto de Histología y Embriología (IHEM)-CONICET, Facultad de Ciencias Médicas, Universidad Nacional de Cuyo, Casilla de Correo 56, Centro Universitario, Parque General San Martín, 5500 Mendoza, Argentina. Phone: 54-261-4205115, ext. 2675. Fax: 54-261-4494117. E-mail for Walter Berón: wberon{at}fmed2.uncu.edu.ar. E-mail for María I. Colombo: mcolombo{at}fmed2.uncu.edu.ar.

Editor: J. T. Barbieri

Infection and Immunity, October 2002, p. 5816-5821, Vol. 70, No. 10
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.10.5816-5821.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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