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Analysis of hepatitis A virus translation in a T7 polymerase-expressing cell line

  • L. E. Whetter
  • S. P. Day
  • E. A. Brown
  • O. Elroy-Stein
  • S. M. Lemon
Part of the Archives of Virology Supplementum book series (ARCHIVES SUPPL, volume 9)

Summary

Hepatitis A virus (HAV) exhibits several characteristics which distinguish it from other picornaviruses, including slow growth in cell culture even after adaptation, and lack of host-cell protein synthesis shut-down. Like other picornaviruses, HAV contains a long 5′ non-translated region (NTR) incorporating an internal ribosomal entry site (IRES), which directs cap-independent translation. We compared HAV IRES-initiated translation with translation initiated by the structurally similar encephalomyocarditis virus (EMCV) IRES, using plasmids in which each of the 5′NTRs is linked inframe with the chloramphenicol acetyltransferase (CAT) gene. Translation was assessed in an HAV- permissive cell line which constitutively expresses T7 RNA polymerase and transcribes high levels of uncapped RNA from these plasmids following transfection. RNAs containing the EMCV IRES were efficiently translated in these cells, while those containing the HAV IRES were translated very poorly. Analysis of translation of these RNAs in the presence of polio virus protein 2A, which shuts down cap-dependent translation, demonstrated that their translation was cap independent. Our results suggest that the HAV IRES may function poorly in these cells, and that inefficient translation may contribute to the exceptionally slow replication cycle characteristic of cell culture-adapted HAV.

Keywords

Internal Ribosomal Entry Site Rabbit Reticulocyte Lysate Internal Ribosomal Entry Site Element Internal Initiation Internal Ribosomal Entry Site Activity 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • L. E. Whetter
    • 1
  • S. P. Day
    • 1
  • E. A. Brown
    • 1
  • O. Elroy-Stein
    • 2
  • S. M. Lemon
    • 1
  1. 1.Department of MedicineThe University of North Carolina at Chapel HillChapel HillUSA
  2. 2.Department of Cell Research and ImmunologyTel Aviv UniversityTel AvivIsrael

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