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Initiation by the yeast viral transcriptase in vitro.

M E Nemeroff, J A Bruenn

    The Journal of Biological Chemistry
    |May 15, 1987
    PubMed
    Summary
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    Yeast viruses possess primer-independent transcriptase activity, initiating RNA transcription without needing pre-existing primers. Unlike higher eucaryotic viruses, they produce uncapped messenger RNAs (mRNAs).

    Area of Science:

    • Virology
    • Molecular Biology
    • Yeast Genetics

    Background:

    • Double-stranded RNA (dsRNA) viruses possess inherent transcriptase activity within their capsids.
    • In viruses like reovirus and yeast viruses, transcription is the initial step in viral replication.
    • Understanding viral transcriptase mechanisms is crucial for comprehending viral replication strategies.

    Purpose of the Study:

    • To investigate the in vitro RNA transcription initiation mechanism of yeast dsRNA viruses.
    • To characterize the 5' terminus of the synthesized plus-strand RNA.
    • To compare the primer dependency and mRNA capping strategies of yeast viruses with those of other eukaryotic dsRNA viruses.

    Main Methods:

    • In vitro transcription assays using purified yeast viral particles.

    Related Experiment Videos

  • Analysis of the 5' end of newly synthesized RNA using enzymatic treatments and chromatography.
  • Comparison of viral particle composition and enzymatic activity.

    • Main Results:

    • Yeast viral transcriptase initiates RNA synthesis in vitro, producing plus-strand RNA with a 5' ppGp terminus.
    • Transcription occurs independently of pre-existing primers.
    • Synthesized messenger RNAs (mRNAs) are uncapped, unlike those from higher eukaryotic dsRNA viruses.
    • Viral particles with a single major capsid polypeptide are transcriptionally and replication-competent.

    Conclusions:

    • Yeast dsRNA viruses exhibit a primer-independent capsid-associated transcriptase activity.
    • The synthesis of uncapped mRNAs distinguishes yeast viruses from their higher eukaryotic counterparts.
    • The yeast viral system offers a simpler model for studying dsRNA virus transcription and replication.