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Volume 91, Issue 7

Murine norovirus-1 3D exhibits RNA-dependent RNA polymerase activity and nucleotidylylates on Tyr of the VPg Free

Abstract

We investigated the roles and biochemical properties of recombinant murine norovirus-1 (MNV-1) 3D in RNA synthesis and virus genome-linked protein (VPg) nucleotidylylation. We therefore expressed VPg and 3D of MNV-1 in . MNV-1 3D exhibited RNA-dependent RNA polymerase (RdRp) activity with poly(A) RNA as a template and MnCl as a cofactor. MNV-1 3D demonstrated optimum RNA-synthesis activity at pH 7.4 and 37 °C in the absence of a primer. Further, VPg was guanylylated by MNV-1 3D in the presence of MnCl in a template-independent manner. The guanylylation reaction conducted with VPg substitution mutants (Y26F, Y40F, Y45F and Y117F) and a deletion mutant (Δ117–124) indicated that Tyr was the probable target site of guanylylation. Homopolymeric RNAs did not enhance VPg guanylylation, whereas -transcribed (−) subgenomic (SG) and (+)SG RNA enhanced VPg guanylylation by 9.2 and 3.2 times, respectively. Within (−)SG RNA, the (−)ORF3 region played a critical role in enhancing VPg guanylylation, suggesting that the MNV-1 ORF3 region of negative-strand RNA contains a -acting element that stimulates 3D-mediated VPg guanylylation.

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2010-07-01
2024-04-27
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Murine norovirus-1 3Dpol exhibits RNA-dependent RNA polymerase activity and nucleotidylylates on Tyr of the VPg
J. Gen. Virol. 91, 1713 (2010); https://doi.org/10.1099/vir.0.020461-0
/content/journal/jgv/10.1099/vir.0.020461-0
/content/journal/jgv/10.1099/vir.0.020461-0
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vol. , part 7, pp. 1713–1722

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