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Related Experiment Videos

Coronavirus polyprotein processing

S R Weiss1, S A Hughes, P J Bonilla

  • 1Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia.

Archives of Virology. Supplementum
|January 1, 1994
PubMed
Summary
This summary is machine-generated.

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Murine hepatitis virus (MHV) gene 1 encodes essential viral proteases and replication machinery. Studies reveal processing differences and potential protease defects in MHV mutants, impacting viral replication.

Area of Science:

  • Virology
  • Molecular Biology
  • Biochemistry

Background:

  • Murine hepatitis virus (MHV) gene 1 is crucial for viral replication and contains two open reading frames (ORFs), ORF 1a and ORF 1b.
  • ORF 1a is predicted to encode multiple protease activities, including papain-like and poliovirus 3C-like enzymes, while ORF 1b may encode polymerase and helicase functions.

Purpose of the Study:

  • To investigate the functional roles of MHV gene 1-encoded proteins, particularly the protease activities within ORF 1a.
  • To characterize the processing of ORF 1a polyproteins and identify potential defects in viral protease activity using MHV mutants.

Main Methods:

  • Detection of ORF 1a and ORF 1b polypeptides in infected cells using antibodies.
  • Analysis of polypeptide processing during cell-free translation versus infected cells.

Related Experiment Videos

  • Characterization of MHV-A59 RNA negative mutants at non-permissive temperatures.

    • Main Results:

    • Only the N-terminal papain-like domain of ORF 1a demonstrated protease activity; other predicted activities remain unconfirmed.
    • Processing of ORF 1a polypeptides differs between cell-free systems and infected cells, suggesting distinct protease involvement.
    • MHV-A59 mutants showed significantly reduced p28 and p65 expression, indicating potential defects in viral protease functions.

    Conclusions:

    • MHV ORF 1a encodes essential viral proteases, with processing complexities influenced by cellular or viral factors.
    • Further investigation into MHV ORF 1b functions, including polymerase and helicase activities, is warranted.
    • MHV protease activity is critical for viral replication, as evidenced by defects in specific mutants.