PMID:21501366

Citation	de Barsy, M, Jamet, A, Filopon, D, Nicolas, C, Laloux, G, Rual, JF, Muller, A, Twizere, JC, Nkengfac, B, Vandenhaute, J, Hill, DE, Salcedo, SP, Gorvel, JP, Letesson, JJ and De Bolle, X (2011) Identification of a Brucella spp. secreted effector specifically interacting with human small GTPase Rab2. Cell. Microbiol. 13:1044-58
Abstract	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-β-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 ΔvirB mutant. In HeLa cells, a ΔricA 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. abortusΔricA 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.
Links	PubMed Online version:10.1111/j.1462-5822.2011.01601.x
Keywords	Animals; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Brucella abortus/pathogenicity; Cell Line; Epithelial Cells/microbiology; Gene Deletion; Host-Pathogen Interactions; Humans; Macrophages/microbiology; Mice; Phagosomes/metabolism; Phagosomes/microbiology; Protein Binding; Protein Interaction Mapping; Two-Hybrid System Techniques; Virulence; Virulence Factors/genetics; Virulence Factors/metabolism; rab2 GTP-Binding Protein/metabolism
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
BRUA2:Q2YQG1		GO:0052050: interaction with host via substance secreted by type IV secretion system	ECO:0000314:		P		Figure 3. TEM-b-lactamase-RicA translocation into RAW264.7 macrophages. Fig. 4. VirB-independent secretion of 3Flag-RicA by B. abortus.	complete CACAO 11199
BRUME:Q8YHR0		GO:0044164: host cell cytosol	ECO:0000314:		C		Fig. 2. Cytosolic localization of HA-tagged RicA	complete CACAO 11200

References

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