f Nucleotide requirements at positions +1 to +4 for the initiation of hepatitis C virus positive-strand RNA synthesis
- Authors: Udvitha Nandasoma1, Christopher McCormick1,†, Stephen Griffin1,‡, Mark Harris1
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1 Institute of Molecular and Cellular Biology, Faculty of Biological Sciences and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK
- Correspondence Mark Harris firstname.lastname@example.org
- First Published Online: 01 May 2011, Journal of General Virology 92: 1082-1086, doi: 10.1099/vir.0.028423-0
- Subject: Animal - RNA
- Issue Published:
RNA virus genome replication requires initiation at the precise terminus of the 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 randomized residues at these positions. There was strong selection for both the wild-type cytosine at position 2, and the wild-type sequence at positions 2–4 (CCU). An adenine residue was well tolerated at positions 3 and 4, which suggests that efficient RNA replication is less dependent on these residues.
Present address: School of Medicine, University of Southampton, Southampton General Hospital, Southampton SO16 6YD, UK.
Present address: Leeds Institute of Molecular Medicine, Faculty of Medicine and Health, University of Leeds, Leeds LS9 7TF, UK.
A supplementary table, showing input and output sequences at the +2, +3 and +4 positions present in the randomized J6/JFH-1 5′ UTR, is available with the online version of this paper.
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