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Structure and expression of the M2 genomic segment of a type 2 killer virus of yeast

E M Hannig, M J Leibowitz

    Nucleic Acids Research
    |June 25, 1985
    PubMed

    Insights

    The M2 double-stranded RNA in Saccharomyces cerevisiae encodes killer toxin and resistance functions. RNA analysis identified a large open reading frame translated into a 28 kd polypeptide, regulating toxin precursor expression.

    Area of Science:

    • Molecular Biology
    • Yeast Genetics
    • Virology

    Background:

    • The M2 double-stranded RNA (dsRNA) species in Saccharomyces cerevisiae confers K2 killer specificity, encoding both toxin and resistance.
    • Understanding the molecular mechanisms of dsRNA gene expression is crucial for yeast killer systems.

    Purpose of the Study:

    • To analyze the M2 dsRNA sequence and identify functional elements involved in toxin precursor translation.
    • To investigate the regulatory mechanisms governing the expression of K2 toxin.

    Main Methods:

    • RNA sequence analysis of M2 dsRNA.
    • In vitro translation of M2 dsRNA heat-cleavage products.
    • Identification of open reading frames and potential regulatory elements.

    Main Results:

    • A large open reading frame was identified on the M2 dsRNA, yielding a 28 kd polypeptide upon in vitro translation.
    • A potential translation initiator AUG triplet within a stem-loop structure near the 5' terminus was found.
    • Putative ribosomal RNA binding sites were identified, suggesting translational regulation of the K2 toxin precursor.

    Conclusions:

    • The M2 dsRNA contains a translatable open reading frame crucial for K2 toxin precursor production.
    • Specific RNA structural features, including a stem-loop and rRNA binding sites, likely regulate translation initiation.
    • Despite limited homology, shared features between M1 and M2 dsRNA suggest conserved mechanisms for gene expression and replication in yeast killer systems.

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