1887

Abstract

Norovirus (NV) 3D is a non-structural protein predicted to play an essential role in the replication of the NV genome. In this study, the characteristics of NV 3D activity and initiation of RNA synthesis have been examined . Recombinant NV 3D, as well as a 3D active-site mutant were expressed in and purified. NV 3D was able to synthesize RNA and displayed flexibility with respect to the use of Mg or Mn as a cofactor. NV 3D yielded two different products when incubated with synthetic RNA : (i) a double-stranded RNA consisting of two single strands of opposite polarity or (ii) the single-stranded RNA template labelled at its 3′ terminus by terminal transferase activity. Initiation of RNA synthesis occurred rather than by back-priming, as evidenced by the fact that the two strands of the double-stranded RNA product could be separated, and by dissociation in time-course analysis of terminal transferase and RNA synthesis activities. In addition, RNA synthesis was not affected by blocking of the 3′ terminus of the RNA template by a chain terminator, sustaining initiation of RNA synthesis. NV 3D displays properties characteristic of RNA-dependent RNA polymerases, allowing the implementation of this enzymic assay for the development and validation of antiviral drugs against NV, a so far non-cultivated virus and an important human pathogen.

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2006-09-01
2019-11-18
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