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Related Experiment Videos

Cold-adapted poliovirus mutants bypass a postentry replication block

A W Dove1, V R Racaniello

  • 1Department of Microbiology, Columbia University College of Physicians & Surgeons, New York, New York 10032, USA.

Journal of Virology
|June 1, 1997
PubMed
Summary
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Poliovirus entry does not require a 135S particle transition. Cold-adapted mutants, selected at 25°C, bypass this step, indicating alternative infection pathways for poliovirus.

Area of Science:

  • Virology
  • Molecular Biology
  • Cell Biology

Background:

  • Poliovirus entry involves a transition from native 160S virions to altered 135S particles.
  • The 135S particle, lacking VP4 and more hydrophobic, is hypothesized to release the viral genome.
  • High particle-to-PFU ratios in poliovirus are linked to nonproductive 135S transitions.

Purpose of the Study:

  • To investigate the role of the 135S particle transition in poliovirus entry and replication.
  • To identify genetic determinants of cold-adapted (ca) poliovirus mutants.
  • To challenge the existing model of poliovirus cellular entry.

Main Methods:

  • Selection and characterization of cold-adapted (ca) poliovirus mutants at 25°C.
  • Analysis of particle conversion to the 135S form.

Related Experiment Videos

  • Site-directed mutagenesis to introduce specific amino acid changes into the wild-type (wt) poliovirus genome.
  • Assessment of viral replication and particle-to-PFU ratios at different temperatures.

    • Main Results:

    • Cold-adapted (ca) mutants replicated at 25°C without converting to 135S particles.
    • The block to wild-type (wt) poliovirus infection at low temperatures is not related to cellular entry.
    • Three specific amino acid changes in the 2C coding region (positions 218, 241, 309) conferred the ca phenotype individually.
    • Sabin vaccine strains and P3/Leon poliovirus also exhibit the ca phenotype.

    Conclusions:

    • The transition to a 135S particle is not an obligatory step for poliovirus entry into cells.
    • Specific mutations in the 2C protein are responsible for the cold-adapted phenotype.
    • The findings necessitate a revision of the current poliovirus entry model.