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• W2062418171 abstract "Coxiella burnetii is an obligate intracellular bacterial pathogen responsible for acute and chronic Q fever. This bacterium harbors a type IV secretion system (T4SS) highly similar to the Dot/Icm of Legionella pneumophila that is believed to be essential for its infectivity. Protein substrates of the Coxiella T4SS are predicted to facilitate the biogenesis of a phagosome permissive for its intracellular growth. However, due to the lack of genetic systems, protein transfer by the C. burnetii Dot/Icm has not been demonstrated. In this study, we report the identification of 32 substrates of the C. burnetii Dot/Icm system using a fluorescence-based β-lactamase (TEM1) translocation assay as well as the calmodulin-dependent adenylate cyclase (CyaA) assay in the surrogate host L. pneumophila . Notably, 26 identified T4SS substrates are hypothetical proteins without predicted function. Candidate secretion substrates were obtained by using ( i ) a genetic screen to identify C. burnetii proteins interacting with DotF, a component of the T4SS, and ( ii ) bioinformatic approaches to retrieve candidate genes that harbor characteristics associated with previously reported substrates of the Dot/Icm system from both C. burnetii and L. pneumophila . Moreover, we have developed a shuttle plasmid that allows the expression of recombinant proteins in C. burnetii as TEM fusion products. Using this system, we demonstrated that a Dot/Icm substrate identified with L. pneumophila was also translocated by C. burnetii in a process that requires its C terminus, providing direct genetic evidence of a functional T4SS in C. burnetii ." @default.
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• W2062418171 date "2010-11-23" @default.
• W2062418171 modified "2023-10-02" @default.
• W2062418171 title "Large-scale identification and translocation of type IV secretion substrates by <i>Coxiella burnetii</i>" @default.
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• W2062418171 doi "https://doi.org/10.1073/pnas.1010485107" @default.
• W2062418171 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/3003115" @default.
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