Nandasoma, Udvitha, McCormick, Chris, Griffin, Stephen D.C. and Harris, Mark
Nucleotide requirements at positions +1 to +4 for the initiation of hepatitis C virus plus (+) strand RNA synthesis
Journal of General Virology (doi:10.1099/vir.0.028423-0). (PMID:21270286).
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RNA virus genome replication requires initiation at the precise terminus of template RNA. To investigate the nucleotide requirements for initiation of hepatitis C virus (HCV) positive strand RNA replication a hammerhead ribozyme was inserted at the 5' end of an HCV subgenomic replicon allowing the generation of replicons with all four possible nucleotides at position 1. This analysis revealed a preference for a purine nucleotide at this position for initiation of RNA replication. The sequence requirements at positions 2-4 in the context of the J6/JFH-1 virus were also examined by selecting replication competent virus from a pool containing randomised residues at these positions. There was a strong selection for both the wildtype C at position 2, and the wildtype sequence at positions 2-4 (CCU) . An A residue was well tolerated at positions 3 and 4, suggesting that efficient RNA replication is less dependent on these residues.
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