DNA Delivery and Genomic Integration into Mammalian Target Cells through Type IV A and B Secretion Systems of Human Pathogens.
DNA Delivery and Genomic Integration into Mammalian Target Cells through Type IV A and B Secretion Systems of Human Pathogens.
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通过人类病原体的IV A和B分泌系统，DNA递送和基因组整合到哺乳动物靶细胞中。
DOI：
10.3389/fmicb.2017.01503
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发表时间：
2017
影响因子：
5.2
通讯作者：
Llosa M
中科院分区：
文献类型：
--
作者：
Guzmán-Herrador DL;Steiner S;Alperi A;González-Prieto C;Roy CR;Llosa M
我们探索了细菌分泌系统作为人类细胞基因组修饰工具的潜力。我们以前表明，外源DNA可以通过人类病原体汉赛巴尔通体的IV A型分泌系统引入人类细胞。此外，递送的DNA共价连接到接合松弛酶TrwC，这促进其整合到受体基因组中。在这项工作中，我们报告说，这种工具可以通过使用不同的松弛酶和分泌系统来适应其他靶细胞。来自质粒RSF 1010的混杂松弛酶MobA可用于以比TrwC更高的效率将DNA递送到人类细胞中。MobA也促进DNA整合，尽管速率低于TrwC。值得注意的是，我们报告说，DNA转移到人类细胞也可以通过两个细胞内的人类病原体，嗜肺军团菌和贝氏柯克斯体，编码一个远亲点/Icm IV型B分泌系统的IV型分泌系统。这表明DNA转移可能是这个分泌系统家族的内在能力，扩大了靶向人类细胞的范围。DNA转移过程的进一步分析表明，通过Dot/Icm募集MobA依赖于IcmSW分子伴侣，这可以解释获得的较高DNA转移速率。最后，我们观察到MobA的存在对C. burnetii，表明干扰Dot/Icm易位的毒力因子。
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.
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影响因子：
3.1
作者：
Celec, Peter;Gardlik, Roman
通讯作者：
Gardlik, Roman
影响因子：
6.7
作者：
Carey KL;Newton HJ;Lührmann A;Roy CR
通讯作者：
Roy CR
影响因子：
3.2
作者：
Fernandez-Gonzalez, Esther;de Paz, Hector D.;Llosa, Matxalen
通讯作者：
Llosa, Matxalen
影响因子：
48
作者：
Gibson, Daniel G.;Young, Lei;Smith, Hamilton O.
通讯作者：
Smith, Hamilton O.
影响因子：
--
作者：
Christie PJ
通讯作者：
Christie PJ