Identification of a Brucella spp. secreted effector specifically interacting with human small GTPase Rab2.

Animals; Bacterial Proteins/genetics/metabolism; Brucella abortus/pathogenicity; Cell Line; Epithelial Cells/microbiology; Gene Deletion; Host-Pathogen Interactions; Humans; Macrophages/microbiology; Mice; Phagosomes/metabolism/microbiology; Protein Binding; Protein Interaction Mapping; Two-Hybrid System Techniques; Virulence; Virulence Factors/genetics/metabolism; rab2 GTP-Binding Protein/metabolism

Abstract :

[en] Bacteria of the Brucella genus are facultative intracellular class III pathogens. These bacteria are able to control the intracellular trafficking of their vacuole, presumably by the use of yet unknown translocated effectors. To identify such effectors, we used a high-throughput yeast two-hybrid screen to identify interactions between putative human phagosomal proteins and predicted Brucella spp. proteins. We identified a specific interaction between the human small GTPase Rab2 and a Brucella spp. protein named RicA. This interaction was confirmed by GST-pull-down with the GDP-bound form of Rab2. A TEM-beta-lactamase-RicA fusion was translocated from Brucella abortus to RAW264.7 macrophages during infection. This translocation was not detectable in a strain deleted for the virB operon, coding for the type IV secretion system. However, RicA secretion in a bacteriological culture was still observed in a DeltavirB mutant. In HeLa cells, a DeltaricA mutant recruits less GTP-locked myc-Rab2 on its Brucella-containing vacuoles, compared with the wild-type strain. We observed altered kinetics of intracellular trafficking and faster proliferation of the B. abortusDeltaricA mutant in HeLa cells, compared with the wild-type control. Altogether, the data reported here suggest RicA as the first reported effector with a proposed function for B. abortus.

Disciplines :

Biochemistry, biophysics & molecular biology

Author, co-author :

de Barsy, Marie

Jamet, Alexandre

Filopon, Didier

Nicolas, Cecile

Laloux, Geraldine

Rual, Jean-Francois

Muller, Alexandre

Twizere, Jean-Claude ; Université de Liège - ULiège > Chimie et bio-industries > Biologie cell. et moléc.

Nkengfac, Bernard

Vandenhaute, Jean

More authors (5 more)

Language :

English

Title :

Identification of a Brucella spp. secreted effector specifically interacting with human small GTPase Rab2.

Publication date :

2011

Journal title :

Cellular Microbiology

ISSN :

1462-5814

eISSN :

1462-5822

Publisher :

Blackwell, Oxford, United Kingdom

Volume :

Issue :

Pages :

1044-58

Peer reviewed :

Peer Reviewed verified by ORBi

Additional URL :

http://onlinelibrary.wiley.com/doi/10.1111/j.1462-5822.2011.01601.x/pdf

Commentary :

Available on ORBi :

since 05 March 2014

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