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Related Experiment Videos

Temperature-dependent host range mutation in vesicular stomatitis virus affecting polypeptide L.

J F Szilágyi, C R Pringle, T M Macpherson

    Journal of Virology
    |May 1, 1977
    PubMed
    Summary
    This summary is machine-generated.

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    Researchers identified the specific protein responsible for a temperature-sensitive mutation in vesicular stomatitis virus (VSV). Dissociation and reconstitution experiments revealed the L polypeptide is affected by the td CE 3 mutation, impacting viral RNA transcription.

    Area of Science:

    • Virology
    • Molecular Biology
    • Genetics

    Background:

    • Vesicular stomatitis virus (VSV) New Jersey, a temperature-dependent host range mutant (td CE 3), exhibits temperature-sensitive RNA transcriptase activity.
    • Understanding the molecular basis of viral mutations is crucial for comprehending viral replication and pathogenesis.

    Purpose of the Study:

    • To determine which specific VSV polypeptide is affected by the td CE 3 mutation.
    • To elucidate the role of viral polypeptides in RNA transcriptase activity.

    Main Methods:

    • Dissociation and reconstitution of viral components from wild-type VSV, the td CE 3 mutant, and its revertant (td CE/R1).
    • Fractionation of viral nucleoprotein preparations using LiCl treatment to isolate L polypeptide (supernatant) and N/NS polypeptides (pellet).

    Related Experiment Videos

  • Assessing RNA transcriptase activity in vitro after reconstituting different supernatant and pellet fractions at permissive (31°C) and restrictive (39°C) temperatures.

    • Main Results:

    • Reconstitution of supernatant and pellet fractions restored significant RNA synthesis at 31°C.
    • At 39°C, RNA synthesis was restored with wild-type and revertant supernatants, but not with the td CE 3 mutant supernatant.
    • The td CE 3 mutant supernatant, when combined with any pellet fraction, showed minimal to no RNA synthesis at the restrictive temperature.

    Conclusions:

    • The L polypeptide is the component affected by the td CE 3 mutation in VSV New Jersey.
    • This mutation specifically impairs the RNA transcriptase activity of the virus at elevated temperatures.