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Related Experiment Videos

Specific Sindbis virus-coded function for minus-strand RNA synthesis

D L Sawicki, S G Sawicki, S Keränen

    Journal of Virology
    |August 1, 1981
    PubMed
    Summary

    Investigating Sindbis virus RNA synthesis revealed distinct temperature-sensitive mutants. Some mutants require specific viral functions for minus-strand RNA synthesis, while others need common components for both plus and minus-strand RNA production.

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    Molecular microbiology·2002

    Area of Science:

    • Virology
    • Molecular Biology
    • Genetics

    Background:

    • Alphavirus replication involves the synthesis of both plus-strand and minus-strand RNA genomes.
    • Temperature-sensitive (ts) mutants are crucial tools for dissecting viral replication steps.
    • Understanding the genetic requirements for RNA synthesis is key to controlling viral infections.

    Purpose of the Study:

    • To investigate the synthesis of Sindbis virus minus-strand RNA using temperature-sensitive mutants.
    • To identify viral functions essential for minus-strand RNA synthesis versus those required for both minus- and plus-strand synthesis.

    Main Methods:

    • Infection of cell cultures with a heat-resistant Sindbis virus strain and complementation groups A, B, F, and G ts mutants.
    • Shifting infected cultures from a permissive (28°C) to a non-permissive (39°C) temperature at 3 hours postinfection.

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  • Monitoring viral RNA synthesis (minus-strand and plus-strand) after temperature shifts.

    • Main Results:

    • Minus-strand RNA synthesis ceased in cultures infected with ts mutants of complementation groups B and F upon shift to 39°C.
    • Minus-strand RNA synthesis continued in cultures infected with parental virus and mutants of complementation groups A and G.
    • In ts11 (group B) infected cultures, minus-strand synthesis ceased while plus-strand synthesis continued; resuming minus-strand synthesis occurred upon return to 28°C.
    • In ts6 (group F) infected cultures, both plus- and minus-strand RNA synthesis were drastically reduced.

    Conclusions:

    • At least one viral function is specifically required for alphavirus minus-strand synthesis, distinct from plus-strand synthesis.
    • A common viral component is necessary for the synthesis of both minus-strand and plus-strand RNAs, as demonstrated by the ts6 mutant.
    • These findings elucidate the complex genetic control of Sindbis virus RNA replication.