A Brucella abortus cstA mutant is defective for association with endoplasmic reticulum exit sites and displays altered trafficking in HeLa cells

Members of the genus Brucella are facultative intracellular pathogenic bacteria able to control maturation of their vacuoles. In several cell types, Brucella is able to reach a proliferation compartment derived from the endoplasmic reticulum (ER). Since ER exit site (ERES) functions are required for...

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Bibliographic Details
Published in:Microbiology (Society for General Microbiology) 2012-10, Vol.158 (Pt 10), p.2610-2618
Main Authors: DE BARSY, Marie, MIRABELLA, Aurélie, LETESSON, Jean-Jacques, DE BOLLE, Xavier
Format: Article
Language:English
Subjects:
Bacterial Proteins - genetics
Bacterial Proteins - metabolism
Bacteriology
Biological and medical sciences
Brucella abortus - metabolism
Brucella abortus - pathogenicity
Endoplasmic Reticulum - metabolism
Epithelial Cells - microbiology
Fundamental and applied biological sciences. Psychology
HeLa Cells
Host-Pathogen Interactions
Humans
Microbiology
Miscellaneous
Mutation
Protein Transport
Proteome
Two-Hybrid System Techniques
Vesicular Transport Proteins - metabolism
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Summary:Members of the genus Brucella are facultative intracellular pathogenic bacteria able to control maturation of their vacuoles. In several cell types, Brucella is able to reach a proliferation compartment derived from the endoplasmic reticulum (ER). Since ER exit site (ERES) functions are required for Brucella proliferation, we performed a yeast two-hybrid screen between human ERES-associated proteins and the predicted brucella proteome. This screening led to the identification of CstA, a conserved protein that specifically interacts with Sec24A, a component of the ERES. We found that a tagged CstA is secreted in Brucella abortus culture medium. This secretion is independent of the type IV secretion system VirB and the flagellum, suggesting that CstA is secreted through another system. We also discovered that a B. abortus cstA mutant is impaired for its association with the Sec23 ERES marker. The B. abortus cstA mutant displayed peculiar trafficking, with reduced association with LAMP1 and Calnexin 12 h post-infection in HeLa cells. However, its intracellular proliferation kinetics was not affected. The data reported here suggest that CstA could be directly or indirectly involved in the control of B. abortus intracellular trafficking in HeLa cells.
ISSN:1350-0872
1465-2080
DOI:10.1099/mic.0.060509-0