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2017 ; 8
(ä): 1503
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DNA Delivery and Genomic Integration into Mammalian Target Cells through Type IV
A and B Secretion Systems of Human Pathogens
#MMPMID28878740
Guzmán-Herrador DL
; Steiner S
; Alperi A
; González-Prieto C
; Roy CR
; Llosa M
Front Microbiol
2017[]; 8
(ä): 1503
PMID28878740
show ga
We explore the potential of bacterial secretion systems as tools for genomic
modification of human cells. We previously showed that foreign DNA can be
introduced into human cells through the Type IV A secretion system of the human
pathogen Bartonella henselae. Moreover, the DNA is delivered covalently attached
to the conjugative relaxase TrwC, which promotes its integration into the
recipient genome. In this work, we report that this tool can be adapted to other
target cells by using different relaxases and secretion systems. The promiscuous
relaxase MobA from plasmid RSF1010 can be used to deliver DNA into human cells
with higher efficiency than TrwC. MobA also promotes DNA integration, albeit at
lower rates than TrwC. Notably, we report that DNA transfer to human cells can
also take place through the Type IV secretion system of two intracellular human
pathogens, Legionella pneumophila and Coxiella burnetii, which code for a
distantly related Dot/Icm Type IV B secretion system. This suggests that DNA
transfer could be an intrinsic ability of this family of secretion systems,
expanding the range of target human cells. Further analysis of the DNA transfer
process showed that recruitment of MobA by Dot/Icm was dependent on the IcmSW
chaperone, which may explain the higher DNA transfer rates obtained. Finally, we
observed that the presence of MobA negatively affected the intracellular
replication of C. burnetii, suggesting an interference with Dot/Icm translocation
of virulence factors.