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Murine leukemia virus mutant with a frameshift in the reverse transcriptase coding region: implications for pol gene

J G Levin, S C Hu, A Rein

    Journal of Virology
    |August 1, 1984
    PubMed
    Summary

    Researchers identified a single base insertion in the pol gene of a B-tropic MuLV mutant, clone 23. This mutation causes premature translation termination, resulting in a truncated reverse transcriptase with reduced activity.

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    Area of Science:

    • Molecular Biology
    • Virology
    • Genetics

    Background:

    • Murine leukemia virus (MuLV) is a retrovirus with a pol gene encoding reverse transcriptase.
    • Clone 23 is a B-tropic MuLV mutant with a defect in its pol gene.
    • Understanding the molecular basis of this mutation is crucial for retroviral research.

    Purpose of the Study:

    • To characterize the molecular defect in the B-tropic MuLV pol mutant, clone 23.
    • To pinpoint the exact mutation within the pol gene responsible for the observed phenotype.
    • To elucidate the genetic organization of the MuLV pol gene.

    Main Methods:

    • Recombinant DNA technology, including gene cloning and subcloning into plasmids (pBR322).
    • Transfection of NIH-3T3 cells with plasmid clones to assess viral properties.

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  • In vivo ligation experiments and sequence analysis of wild-type and mutant pol genes.

    • Main Results:

    • A plasmid clone, pRTM, successfully reproduced the properties of clone 23 virus-infected cells.
    • The mutation was localized to a 400-base-pair region within the pol gene.
    • A single base (cytosine) insertion was identified, causing a frameshift and premature termination of translation.

    Conclusions:

    • The molecular defect in clone 23 is a single-base insertion in the pol gene, leading to a truncated reverse transcriptase.
    • This mutation results in significantly reduced enzymatic activity of the polymerase.
    • The findings contribute to understanding the genetic organization and functional domains of the MuLV pol gene.