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  • The EMBO Journal (2004) 23, 4072 - 4081
  • doi:10.1038/sj.emboj.7600408

Published online: 16 September 2004

Cell-line-induced mutation of the rotavirus genome alters expression of an IRF3-interacting protein

Karen Kearneya, Dayue Chenb, Zenobia F Taraporewala, Patrice Vendec, Yasutaka Hoshino, Maria Alejandra Tortorici, Mario Barro and John T Patton

  1. Laboratory of Infectious Diseases, National Institutes of Allergy and Infectious Diseases, Bethesda, MD, USA

Correspondence to:

John T Patton, Laboratory of Infectious Diseases, National Institutes of Allergy and Infectious Diseases, 50 South Drive MSC 8026, NIH, Bethesda, MD 20892, USA. Tel.: +1 301 496 5227; Fax: +1 301 496 8312; E-mail: jpatton@niaid.nih.gov

aPresent address: Department of Microbiology, University College Cork, Cork, Ireland

bPresent address: Infectious Diseases Research, Eli Lilly and Company, Lilly Research Laboratories, Indianapolis, IN 46285, USA

cPresent address: Virologie Moléculaire et Structurale, Unité Mixte de Recherche, CNRS-INRA, 91198 Gif-sur-Yvette, France

Received 7 July 2004; Accepted 18 August 2004

Rotavirus, a cause of severe gastroenteritis, contains a segmented double-stranded (ds)RNA genome that replicates using viral mRNAs as templates. The highly conserved 3'-consensus sequence (3'CS), UGUGACC, of the mRNAs promotes dsRNA synthesis and enhances translation. We have found that the 3'CS of the gene (g5) encoding NSP1, an antagonist of interferon signaling, undergoes rapid mutation when rhesus rotavirus (RRV) is serially passaged at high multiplicity of infection (MOI) in cells permitting high titer growth. These mutations increase the promoter activity of the g5 3'-sequence, but decrease its activity as a translation enhancer. The location of the mutations defines the minimal essential promoter for dsRNA synthesis as URN0–5CC. Under passage conditions where cell-to-cell spread of the virus is required to complete infection (low MOI), the 3'CS is retained due to the need for NSP1 to be expressed at levels sufficient to prevent establishment of the antiviral state. These data demonstrate that host cell type and propagation conditions affect the capacity of RRV to produce the virulence gene product NSP1, an important consideration in producing RRV-based vaccines.

  • Keywords:

    • gene expression,
    • interferon regulatory factor 3,
    • RNA replication,
    • rotavirus,
    • vaccine production