Incorporation of Plasmid DNA Into Bacterial Membrane Vesicles by Peptidoglycan Defects in Escherichia coli.
Incorporation of Plasmid DNA Into Bacterial Membrane Vesicles by Peptidoglycan Defects in Escherichia coli.
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DOI：
10.3389/fmicb.2021.747606
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发表时间：
2021
影响因子：
5.2
通讯作者：
Tashiro Y
中科院分区：
文献类型：
--
作者：
Aktar S;Okamoto Y;Ueno S;Tahara YO;Imaizumi M;Shintani M;Miyata M;Futamata H;Nojiri H;Tashiro Y
膜囊泡（MV）由各种原核生物释放，并在递送各种细胞-细胞相互作用因子中发挥作用。最近的研究已经确定这些囊泡能够作为水平基因转移的介质发挥作用。外膜囊泡（OMV）是一种由革兰氏阴性菌释放的MV，主要由外膜和周质成分组成;然而，为什么DNA包含在OMV中仍然是未知的。我们的研究旨在了解革兰氏阴性菌中定位于细胞质中的DNA并入OMV的机制。我们使用携带非接合、非移动和高拷贝质粒pUC 19及其包括Δ nlpI、Δ rseA和Δ tolA的超囊泡突变体的大肠杆菌BW 25113细胞比较了与OMV相关的DNA。在来自Δ nlpI的OMV中，每个囊泡的质粒拷贝数增加，其中肽聚糖（PG）的分解和合成发生改变。当补充1%甘氨酸抑制PG合成，OMV形成和质粒拷贝每囊泡中的野生型增加。细菌膜条件试验表明，在对数生长期后期，甘氨酸的存在增加了细菌膜的通透性，而在对数生长期后期，没有发生细胞裂解。此外，速冻深蚀刻和复制电子显微镜观察显示，外内膜囊泡（O-IMVs）的存在下形成的甘氨酸。因此，建议在PG损伤条件下将DNA掺入OMV的两种建议途径。这些途径包括由于增加的膜渗透和O-IMV形成而导致的DNA泄漏。此外，我们的研究结果有助于更好地理解自然界中发生的囊泡介导的水平基因转移和利用MV的DNA货物。
Membrane vesicles (MVs) are released by various prokaryotes and play a role in the delivery of various cell-cell interaction factors. Recent studies have determined that these vesicles are capable of functioning as mediators of horizontal gene transfer. Outer membrane vesicles (OMVs) are a type of MV that is released by Gram-negative bacteria and primarily composed of outer membrane and periplasm components; however, it remains largely unknown why DNA is contained within OMVs. Our study aimed to understand the mechanism by which DNA that is localized in the cytoplasm is incorporated into OMVs in Gram-negative bacteria. We compared DNA associated with OMVs using Escherichia coli BW25113 cells harboring the non-conjugative, non-mobilized, and high-copy plasmid pUC19 and its hypervesiculating mutants that included ΔnlpI, ΔrseA, and ΔtolA. Plasmid copy per vesicle was increased in OMVs derived from ΔnlpI, in which peptidoglycan (PG) breakdown and synthesis are altered. When supplemented with 1% glycine to inhibit PG synthesis, both OMV formation and plasmid copy per vesicle were increased in the wild type. The bacterial membrane condition test indicated that membrane permeability was increased in the presence of glycine at the late exponential phase, in which cell lysis did not occur. Additionally, quick-freeze deep-etch and replica electron microscopy observations revealed that outer-inner membrane vesicles (O-IMVs) are formed in the presence of glycine. Thus, two proposed routes for DNA incorporation into OMVs under PG-damaged conditions are suggested. These routes include DNA leakage due to increased membrane permeation and O-IMV formation. Additionally, our findings contribute to a greater understanding of the vesicle-mediated horizontal gene transfer that occurs in nature and the utilization of MVs for DNA cargo.
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影响因子：
4.4
作者：
Klieve, AV;Yokoyama, MT;Mawhinney, EL
通讯作者：
Mawhinney, EL
影响因子：
4.8
作者：
BOUVERET, E;DEROUICHE, R;BENEDETTI, H
通讯作者：
BENEDETTI, H
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作者：
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影响因子：
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作者：
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通讯作者：
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