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Synthesis of alpha (immediate-early) proteins in Vero cells infected with pseudorabies virus

M L Fenwick, M McMenamin

    The Journal of General Virology
    |September 1, 1984
    PubMed

    Insights

    Pseudorabies virus immediate-early protein synthesis was investigated. Three distinct alpha polypeptides (A, B, C) were identified in infected Vero cells, with protein A being the primary immediate-early protein.

    Area of Science:

    • Virology
    • Molecular Biology
    • Cell Biology

    Background:

    • Pseudorabies virus (PRV) is an alphaherpesvirus that infects various animal species.
    • Understanding the synthesis and function of viral immediate-early (IE) proteins is crucial for comprehending viral replication strategies.
    • IE proteins play a key role in initiating the viral life cycle and modulating host cell functions.

    Purpose of the Study:

    • To investigate the synthesis and characteristics of alpha (immediate-early) polypeptides in Vero cells infected with pseudorabies virus.
    • To elucidate the relationship between different synthesized viral proteins and their potential roles during infection.
    • To compare the synthesized proteins with known immediate-early proteins of other herpesviruses.

    Main Methods:

    • Vero cells were infected with pseudorabies virus and treated with cycloheximide to accumulate alpha mRNA.
    • Accumulated mRNA was translated in vivo (with actinomycin D) and in vitro.
    • Proteins were analyzed by electrophoresis, phosphorylation assays, chymotrypsin digestion, and subcellular fractionation.

    Main Results:

    • Three distinct virus-specific proteins (A, B, C) with molecular weights of 180,000, 190,000, and 200,000 Da were synthesized in intact cells.
    • Only protein A was detected during in vitro translation; proteins B and C were not found in normally infected cells.
    • Proteins A and B were phosphorylated, associated with the nuclear fraction, and showed similar chymotrypsin digestion patterns, suggesting structural similarity.

    Conclusions:

    • Protein A is presumed to be the single immediate-early protein, analogous to ICP 4 of herpes simplex virus.
    • The functions of proteins B and C remain unknown, but they may represent intermediate stages in the synthesis or transport of protein A.
    • The progression of protein synthesis might depend on an unstable cellular protein, as indicated by cycloheximide treatment effects.

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