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Related Experiment Video

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Live Cell Imaging of Alphaherpes Virus Anterograde Transport and Spread
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Imaging poliovirus entry in live cells.

Boerries Brandenburg1, Lily Y Lee, Melike Lakadamyali

  • 1Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, United States of America.

Plos Biology
|July 12, 2007
PubMed
Summary
This summary is machine-generated.

Poliovirus rapidly releases its RNA genome after entering cells via an endocytic mechanism, not by directly breaching the plasma membrane. This efficient genome delivery is key to poliovirus infection.

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Area of Science:

  • Virology
  • Cell Biology
  • Molecular Biology

Background:

  • Mechanisms of animal virus entry, particularly for nonenveloped viruses, remain poorly understood.
  • Technical challenges in visualizing viral gene delivery and high particle-to-plaque forming unit ratios complicate studies.
  • Distinguishing productive from nonproductive viral pathways is difficult.

Purpose of the Study:

  • To characterize the early events of poliovirus (PV) infection, focusing on genome delivery.
  • To investigate the role of cell trafficking pathways in viral RNA release.
  • To resolve the debate on whether PV breaches the plasma membrane or uses endocytosis.

Main Methods:

  • Combined live-cell imaging tracking viral capsid and genome with infectivity-based RNA release assays.
  • Systematically probed effects of cell trafficking inhibitors on RNA genome delivery.
  • Utilized high-resolution imaging to observe viral particle internalization and genome release.

Main Results:

  • Poliovirus genome release is highly efficient and rapid, not limiting overall infectivity.
  • PV enters cells via an endocytic mechanism independent of clathrin, caveolin, flotillin, and microtubules.
  • Genome release occurs immediately after internalization from vesicles or membrane invaginations near the plasma membrane.
  • Entry is dependent on tyrosine kinase and actin.

Conclusions:

  • Poliovirus utilizes an endocytic pathway for genome delivery, not direct plasma membrane penetration.
  • Genome release is a rapid and efficient process following internalization.
  • The developed imaging assay is broadly applicable for studying nonenveloped virus entry mechanisms.