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The Encapsulation of Cell-free Transcription and Translation Machinery in Vesicles for the Construction of Cellular Mimics
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The Encapsulation of Cell-free Transcription and Translation Machinery in Vesicles for the Construction of Cellular Mimics

Published on: October 21, 2013

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Bacterial membrane vesicles transport their DNA cargo into host cells

Natalie J Bitto1, Ross Chapman1, Sacha Pidot2

  • 1Centre for Innate Immunity and Infectious Diseases, Hudson Institute of Medical Research and the Department of Molecular and Translational Sciences, Monash University, Clayton, Victoria, 3168, Australia.

Scientific Reports
|August 3, 2017
PubMed

Insights

Bacterial outer membrane vesicles (OMVs) package chromosomal DNA, with internal DNA enriched in virulence and resistance genes. These OMVs deliver DNA into host cells, impacting bacterial interactions and vaccine development.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Bacterial outer membrane vesicles (OMVs) are released by bacteria and contain various molecules.
  • While OMVs are known to carry proteins and other components, their DNA content and function are poorly understood.
  • Previous studies suggested DNA presence in OMVs, but its nature and host cell interaction capabilities were unclear.

Purpose of the Study:

  • To investigate the presence, nature, and intracellular delivery of chromosomal DNA within bacterial OMVs.
  • To determine if OMV-associated DNA can be transported into eukaryotic host cells.
  • To explore the potential role of OMV-DNA in bacterial-host interactions and its implications for vaccine strategies.

Main Methods:

  • Characterization of DNA content in OMVs shed by bacteria during exponential growth.
  • Analysis of DNA localization (external vs. internal) within OMVs.
  • Genomic analysis of internally packaged DNA to identify enriched chromosomal regions.
  • Demonstration of OMV-mediated DNA transfer into eukaryotic cells using PCR detection in the nuclear fraction.

Main Results:

  • Bacterial chromosomal DNA is packaged into OMVs during exponential growth.
  • A significant portion of OMV DNA is located on the external surface, with a smaller fraction internally packaged.
  • Internally located DNA within Pseudomonas aeruginosa OMVs is enriched for genes related to virulence, stress response, antibiotic resistance, and metabolism.
  • OMVs successfully deliver their DNA cargo into eukaryotic cells, with detectable DNA in the host cell nuclear fraction.

Conclusions:

  • Bacterial OMVs actively package and transport chromosomal DNA into host cells.
  • The enrichment of specific gene sets within internal OMV DNA suggests a selective packaging mechanism.
  • OMV-associated DNA plays a role in bacterial-host cell communication and interaction.
  • These findings have significant implications for understanding bacterial pathogenesis and developing OMV-based vaccines.

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