Article (Scientific journals)
Identification of a Brucella spp. secreted effector specifically interacting with human small GTPase Rab2.
de Barsy, Marie; Jamet, Alexandre; Filopon, Didier et al.
2011In Cellular Microbiology, 13 (7), p. 1044-58
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Keywords :
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
Hill, David E.
Salcedo, Suzana P.
Gorvel, Jean-Pierre
Letesson, Jean-Jacques
De Bolle, Xavier
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Title :
Identification of a Brucella spp. secreted effector specifically interacting with human small GTPase Rab2.
Publication date :
Journal title :
Cellular Microbiology
Publisher :
Blackwell, Oxford, United Kingdom
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Peer reviewed :
Peer Reviewed verified by ORBi
Commentary :
(c) 2011 Blackwell Publishing Ltd.
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