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囊泡保护的细胞外DNA对嗜热栖热菌属水平基因转移的贡献

Contribution of vesicle-protected extracellular DNA to horizontal gene transfer in Thermus spp.

作者信息

Blesa Alba, Berenguer José

机构信息

Center of Molecular Biology Severo Ochoa, Autonomous University of Madrid-CSIC, Madrid, Spain.

出版信息

Int Microbiol. 2015 Sep;18(3):177-87. doi: 10.2436/20.1501.01.248.

Abstract

Highly efficient apparatus for natural competence and conjugation have been shown as the major contributors to horizontal gene transfer (HGT) in Thermus thermophilus. In practical terms, both mechanisms can be distinguished by the sensitivity of the former to the presence of DNAse, and the requirement for cell to cell contacts in the second. Here we demonstrate that culture supernatants of different strains of Thermus spp. produce DNAse-resistant extracellular DNA (eDNA) in a growth-rate dependent manner. This eDNA was double stranded, similar in size to isolated genomic DNA (around 20 kbp), and represented the whole genome of the producer strain. Protection against DNAse was the consequence of association of the eDNA to membrane vesicles which composition was shown to include a great diversity of cell envelope proteins with minor content of cytoplasmic proteins. Access of the recipient cell to the protected eDNA depended on the natural competence apparatus and elicited the DNA-DNA interference defence mediated by the Argonaute protein. We hypothesize on the lytic origin of the eDNA carrying vesicles and discuss the relevance of this alternative mechanism for HGT in natural thermal environments.

摘要

高效的自然感受态和接合装置已被证明是嗜热栖热菌中水平基因转移(HGT)的主要促成因素。实际上,这两种机制可以通过前者对DNA酶存在的敏感性以及后者对细胞间接触的需求来区分。在这里,我们证明了不同嗜热栖热菌菌株的培养上清液以生长速率依赖的方式产生抗DNA酶的细胞外DNA(eDNA)。这种eDNA是双链的,大小与分离的基因组DNA相似(约20 kbp),代表了产生菌株的整个基因组。对DNA酶的抗性是eDNA与膜泡结合的结果,膜泡的组成显示包括多种细胞包膜蛋白和少量细胞质蛋白。受体细胞对受保护的eDNA的获取取决于自然感受态装置,并引发由Argonaute蛋白介导的DNA-DNA干扰防御。我们推测携带eDNA的囊泡的裂解起源,并讨论这种替代机制在自然热环境中对HGT的相关性。

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