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Bacterial type IV secretion systems: a tool for DNA transfer into mammalian cells

AutorFernández-González, Esther ; De Paz, Héctor D. ; Dehio, Christoph; Sangari, Félix J. ; Llosa, Matxalen
Fecha de publicación2009
CitaciónNew Biotechnology 25S: S35 (2009)
ResumenType IV secretion systems (T4SS) are versatile machineries for the selective secretion of macromolecules that are involved in different biological processes such as pathogenicity (pT4SS) or horizontal DNA transfer by conjugation (cT4SS). Human intracellular pathogens, such a Bartonella henselae and Brucella suis, use a pT4SS to deliver effector proteins into the cytoplasm of eukaryotic cells. To recruit its specific substrate each T4SS has a coupling protein (CP). We have previously shown that CP can interact with heterologous T4SS.We investigated if a pT4SS could be used to deliver DNA (the substrate of the cT4SS) into its target mammalian cells. To test if the pT4SS of Bartonella and Brucella are able to recruit the cT4SS substrate of the conjugative plasmid R388, we constructed a plasmid coding for the R388 components required for substrate recruitment and a GFP cassette that is expressed only in the eukaryotic cells. Mammalian cells were infected with bacteria harbouring the plasmid. DNA transfer from intracellular bacteria is detected by GFP expression using flow cytometry. In the case of Bartonella about 1% of cells were GFP positive, indicating DNA mobilization from the bacteria into the human cells. The transfer was dependent on the T4SS of the bacteria and the CP of the plasmid, confirming that we have achieved the recruitment of a heterologous DNA substrate by a pT4SS. This system will be an interesting tool for the introduction of any DNA molecule into specific cellular types, since each pathogen containing a T4SS has a different target tissue. This has prompted us to test DNA transfer through pT4SS of other human pathogens. Experiments with Brucella are underway.
DescripciónTrabajo presentado al 14th European Congress on Biotechnology celebrado en Barcelona del 13 al 16 de septiembre de 2009.
Identificadoresdoi: 10.1016/j.nbt.2009.06.086
issn: 1871-6784
e-issn: 1876-4347
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