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

Marie de Barsy, Alexandre Jamet, Didier Filopon, Cécile Nicolas, Géraldine Laloux, Jean-François Rual, Alexandre Muller, Jean-Claude Twizere, Bernard Nkengfac, Jean Vandenhaute, David E Hill, Suzana P Salcedo, Jean-Pierre Gorvel, Jean-Jacques Letesson, Xavier De Bolle

Résultats de recherche: Contribution à un journal/une revueArticle

Résumé

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.
langue originaleAnglais
Pages (de - à)1044-58
Nombre de pages15
journalCellular microbiology
Volume13
Numéro de publication7
Les DOIs
étatPublié - 1 juil. 2011

Empreinte digitale

Brucella
Monomeric GTP-Binding Proteins
Brucella abortus
Vacuoles
HeLa Cells
Bacteria
Proteins
Operon
Guanosine Triphosphate
Yeasts
Macrophages
Infection

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de Barsy, Marie ; Jamet, Alexandre ; Filopon, Didier ; Nicolas, Cécile ; Laloux, Géraldine ; Rual, Jean-François ; Muller, Alexandre ; Twizere, Jean-Claude ; Nkengfac, Bernard ; Vandenhaute, Jean ; Hill, David E ; Salcedo, Suzana P ; Gorvel, Jean-Pierre ; Letesson, Jean-Jacques ; De Bolle, Xavier. / Identification of a Brucella spp. secreted effector specifically interacting with human small GTPase Rab2. Dans: Cellular microbiology. 2011 ; Vol 13, Numéro 7. p. 1044-58.
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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.",
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Identification of a Brucella spp. secreted effector specifically interacting with human small GTPase Rab2. / de Barsy, Marie; Jamet, Alexandre; Filopon, Didier; Nicolas, Cécile; Laloux, Géraldine; Rual, Jean-François; Muller, Alexandre; Twizere, Jean-Claude; Nkengfac, Bernard; Vandenhaute, Jean; Hill, David E; Salcedo, Suzana P; Gorvel, Jean-Pierre; Letesson, Jean-Jacques; De Bolle, Xavier.

Dans: Cellular microbiology, Vol 13, Numéro 7, 01.07.2011, p. 1044-58.

Résultats de recherche: Contribution à un journal/une revueArticle

TY - JOUR

T1 - Identification of a Brucella spp. secreted effector specifically interacting with human small GTPase Rab2

AU - de Barsy, Marie

AU - Jamet, Alexandre

AU - Filopon, Didier

AU - Nicolas, Cécile

AU - Laloux, Géraldine

AU - Rual, Jean-François

AU - Muller, Alexandre

AU - Twizere, Jean-Claude

AU - Nkengfac, Bernard

AU - Vandenhaute, Jean

AU - Hill, David E

AU - Salcedo, Suzana P

AU - Gorvel, Jean-Pierre

AU - Letesson, Jean-Jacques

AU - De Bolle, Xavier

N1 - © 2011 Blackwell Publishing Ltd.

PY - 2011/7/1

Y1 - 2011/7/1

N2 - 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.

AB - 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.

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