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Coronavirus JHM: nucleotide sequence of the mRNA that encodes nucleocapsid protein

M A Skinner, S G Siddell

    Nucleic Acids Research
    |August 11, 1983
    PubMed

    Insights

    Researchers cloned a DNA copy of the Mouse Hepatitis Virus JHM nucleocapsid protein mRNA. This clone specifically inhibited nucleocapsid protein synthesis in vitro, aiding in sequence determination and functional analysis.

    Area of Science:

    • Molecular Biology
    • Virology
    • Genetics

    Background:

    • Mouse Hepatitis Virus (MHV) JHM is a significant pathogen.
    • Understanding viral protein function is crucial for developing antivirals.
    • The nucleocapsid (N) protein plays a key role in viral replication.

    Purpose of the Study:

    • To clone and sequence the mRNA encoding the MHV JHM nucleocapsid protein.
    • To analyze the predicted protein sequence for functional domains.
    • To investigate potential roles of sequence features in viral gene expression.

    Main Methods:

    • Cloning of mRNA-derived DNA into pAT153 vector.
    • In vitro synthesis inhibition assays.
    • Subcloning into M13 vectors for sequencing.
    • Chain-terminator sequencing method.
    • Bioinformatic analysis of the DNA sequence.

    Main Results:

    • A DNA copy of the nucleocapsid protein mRNA was successfully cloned.
    • The cloned DNA specifically inhibited in vitro nucleocapsid protein synthesis.
    • The complete 1767-base sequence, including a poly (A) tail, was determined.
    • An open-reading frame predicted a basic protein (mol.wt. 49700) with 5 basic regions and a serine-rich area.
    • The 5' non-coding region contained a triple repeat (AATCT), potentially involved in subgenomic mRNA production.

    Conclusions:

    • The determined sequence provides a detailed molecular map of the MHV JHM nucleocapsid protein.
    • The identified structural features, such as basic and serine-rich regions, may be functionally significant for viral replication and phosphorylation.
    • The 5' non-coding sequence feature suggests a regulatory role in viral gene expression.

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