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• W2102470405 abstract "RNA virus polymerases must initiate replicative RNA synthesis with extremely high accuracy to maintain their genome termini and to avoid generating defective genomes. For the single-stranded negative-sense RNA viruses, it is not known how this accuracy is achieved. To investigate this question, mutations were introduced into the 3′ terminal base of a respiratory syncytial virus (RSV) template, and the RNA products were examined to determine the impact of the mutation. To perform the assay, RNA replication was reconstituted using a modified minireplicon system in which replication was limited to a single step. Importantly, this system allowed analysis of RSV RNA generated intracellularly, but from a defined template that was not subject to selection by replication. Sequence analysis of RNA products generated from templates containing 1U-C and 1U-A substitutions showed that, in both cases, replication products were initiated with a nontemplated, WT A residue, rather than a templated G or U residue, indicating that the polymerase selects the terminal NTP independently of the template. Examination of a template in which the position 1 nucleotide was deleted supported these findings. This mutant directed efficient replication at ∼60% of WT levels, and its product was found to be initiated at the WT position (−1 relative to the template) with a WT A residue. These findings show that the RSV replicase selects ATP and initiates at the correct position, independently of the first nucleotide of the template, suggesting a mechanism by which highly accurate replication initiation is achieved." @default.
• W2102470405 created "2016-06-24" @default.
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• W2102470405 date "2010-05-17" @default.
• W2102470405 modified "2023-10-04" @default.
• W2102470405 title "Evidence that the polymerase of respiratory syncytial virus initiates RNA replication in a nontemplated fashion" @default.
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• W2102470405 doi "https://doi.org/10.1073/pnas.0913065107" @default.
• W2102470405 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/2890450" @default.
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