Coronaviruses pp 221-226 | Cite as

A Newly Identified MHV-A59 ORF1a Polypeptide p65 is Temperature Sensitive in Two RNA Negative Mutants

  • Scott A. Hughes
  • Mark R. Denison
  • Pedro Bonilla
  • Julian L. Leibowitz
  • Ralph S. Baric
  • Susan R. Weiss
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 342)


Polypeptide products of MHV-A59 gene 1 have been identified in infected DBT cells and in the products of in vitro translations of genome RNA (1,2). In this paper we report the identification in infected cell lysates of a 65-kDa polypeptide (p65) encoded in ORF la. Studies on the kinetics of appearance and processing of p65 show that p65 is detectable after p28 but before the appearance of p290, p240 and p50. No homologue of the p65 polypeptide identified in infected cell lysates was immunoprecipitated from in vitro translations of genomic RNA, providing further evidence that in vitro processing of polypeptides encoded in ORF la of gene 1 differs from that which occurs late in infection of DBT cells. Although the function of p65 is unknown, two MHV-A59 ts mutants isolated and characterized by Baric et al. (3,4) do not produce detectable levels of p65 at the non-permissive temperature indicating that p65 may play an important role in the virus life cycle.


Permissive Temperature Chase Period Pulse Chase Infected Cell Lysate Papain Protease 
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Copyright information

© Springer Science+Business Media New York 1994

Authors and Affiliations

  • Scott A. Hughes
    • 1
  • Mark R. Denison
    • 2
  • Pedro Bonilla
    • 1
  • Julian L. Leibowitz
    • 3
  • Ralph S. Baric
    • 4
  • Susan R. Weiss
    • 1
  1. 1.University of Pennsylvania School of MedicinePhila.USA
  2. 2.Vanderbilt University Medical CenterNashvilleUSA
  3. 3.University of TexasHoustonUSA
  4. 4.UNC School of Public HealthChapel HillUSA

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