論文

査読有り 国際誌
2020年8月14日

Multilamellar and multivesicular outer membrane vesicles produced by a Buttiauxella agrestis tolB mutant.

Applied and environmental microbiology
  • Kotaro Takaki
  • ,
  • Yuhei O Tahara
  • ,
  • Nao Nakamichi
  • ,
  • Yusuke Hasegawa
  • ,
  • Masaki Shintani
  • ,
  • Moriya Ohkuma
  • ,
  • Makoto Miyata
  • ,
  • Hiroyuki Futamata
  • ,
  • Yosuke Tashiro

AEM.01131-20
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1128/AEM.01131-20

Outer membrane vesicles (OMVs) are naturally released from gram-negative bacteria and play important roles in various biological functions. Released vesicles are not uniform in shape, size, and characteristics, and little is known about this diversity of OMVs. Here, we show that deletion of tolB, which encodes a part of the Tol-Pal system, leads to the production of multiple types of vesicles and increases overall vesicle production in the high-vesicle-forming strain Buttiauxella agrestis JCM 1090T The ΔtolB mutant produced small OMVs and multilamellar/multivesicular OMVs (M-OMVs), as well as vesicles with a strikingly similarity to the wild type. M-OMVs, previously undescribed, contained triple lamellar membrane vesicles and multiple vesicle-incorporating vesicles. Ultracentrifugation enabled the separation and purification of each type of OMVs released from the ΔtolB mutant, and visualization by quick-freeze deep-etch and replica electron microscopy indicated that M-OMVs are composed of several lamellar membranes. Visualization of intracellular compartments of ΔtolB cells showed that vesicles were accumulated in the broad periplasm, which is probably due to the low linkage between the outer and inner membranes attributed to the Tol-Pal defect. The outer membrane was invaginating inward by wrapping a vesicle and the precursor of M-OMVs existed in the cell. Thus, we demonstrated a novel type of bacterial OMVs and showed that unconventional processes enable B. agrestis ΔtolB mutant to form unique vesicles.IMPORTANCE Membrane vesicle (MV) formation has been recognized as a common mechanism in prokaryotes, and MVs play critical roles in intercellular interaction. However, a broad range of MV types and their multiple production processes make it difficult to gain a comprehensive understanding of MVs. In this work, using vesicle separation and electron microscopic analyses, we demonstrated that diverse types of outer membrane vesicles (OMVs) were released from an engineered strain, Buttiauxella agrestis 1090T ΔtolB mutant. We also discovered a previously undiscovered type of vesicles, multilamellar/multivesicular outer membrane vesicles (M-OMVs), which were released by this mutant using an unconventional process. These findings have facilitated considerable progress in understanding MV diversity and expanding the utility of MVs in biotechnological applications.

リンク情報
DOI
https://doi.org/10.1128/AEM.01131-20
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/32801184
URL
https://aem.asm.org/content/early/2020/08/11/AEM.01131-20/article-info
ID情報
  • DOI : 10.1128/AEM.01131-20
  • PubMed ID : 32801184

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