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Temperature-sensitive simian virus 40 mutant defective in a late function.

S Kit, S Tokuno, K Nakajima

    Journal of Virology
    |September 1, 1970
    PubMed
    Summary
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    A new temperature-sensitive simian virus 40 (SV40) mutant, tsTNG-1, shows a conditional lethal mutation in a late viral gene. This SV40 mutant replicates poorly at high temperatures but can still transform cells and enhance DNA synthesis.

    Area of Science:

    • Virology
    • Molecular Biology
    • Genetics

    Background:

    • Simian virus 40 (SV40) is a versatile DNA tumor virus used extensively in molecular biology research.
    • Temperature-sensitive mutants are valuable tools for studying viral gene function and replication cycles.

    Purpose of the Study:

    • To isolate and characterize a novel temperature-sensitive mutant of SV40.
    • To investigate the nature of the mutation and its effects on viral replication and gene expression.

    Main Methods:

    • Isolation of a temperature-sensitive mutant (tsTNG-1) from nitrosoguanidine-treated SV40-infected monkey kidney cells.
    • Replication studies at permissive (33.5 C) and non-permissive (38.7 C) temperatures.
    • Temperature-shift experiments to determine the timing of gene function block.

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  • Analysis of viral DNA synthesis, antigen production, and cellular effects (DNA synthesis, transformation).

    • Main Results:

    • SV40tsTNG-1 exhibited a 3,000-fold reduction in replication at 38.7 C compared to 33.5 C.
    • Plaque formation was significantly inhibited at the non-permissive temperature.
    • Temperature-shift experiments indicated a block in a late viral gene function.
    • The mutant synthesized T antigen, capsid antigens, viral DNA, and induced thymidine kinase activity at both temperatures.
    • SV40tsTNG-1 enhanced cellular DNA synthesis and transformed mouse kidney cells.

    Conclusions:

    • SV40tsTNG-1 harbors a conditional lethal mutation affecting a late viral gene function.
    • The mutant retains the ability to induce T antigen, synthesize viral DNA, and transform cells.
    • This mutant provides a valuable tool for dissecting late-stage SV40 gene functions.