{"doi":"10.1101/2020.02.12.946558","title":"Induced copy-back RNA synthesis as a novel therapeutic mechanism against RNA viruses","abstract":"The viral RNA-dependent RNA polymerase (RdRp) is a well-established target for development of broad-spectrum antiviral therapeutics. Incorporation of ribonucleotide analogues by the RdRp will either cause termination of RNA synthesis or mutagenesis of the RNA product. We demonstrated recently that incorporation of a pyrazine-carboxamide ribonucleotide into nascent RNA leads to pausing and backtracking of the elongating RdRp. Here, we provide evidence for the single-stranded RNA product of backtracking serving as an intermediate in RdRp-catalyzed, template-switching reactions. This intermediate is used for both intramolecular template-switching (copy-back RNA synthesis) and intermolecular template-switching (homologous RNA recombination). The use of a magnetic-tweezers platform to monitor RdRp elongation dynamics permitted direct observation of copy-back synthesis and illuminated properties of the RdRp that promote copy-back synthesis, including stability of the RdRp-nascent-RNA complex and the dimensions of the RdRp nucleic-acid-binding channel. In cells, recombination was stimulated by the presence of a pyrazine-carboxamide ribonucleotide. The effect of the drug on recombination was diminished for a recombination-defective virus, but this virus was not resistant to the drug. The discovery that a ribonucleotide analogue can induce copy-back RNA synthesis suggests that this third mechanistic class of compounds may function by promoting formation of defective viral genomes. This study identifies RdRp-catalyzed intra- and intermolecular template switching as a viable new mechanistic target with potentially broad-spectrum appeal.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2020,"id":120850,"datarank":0.32958368660043297,"base_score":2.1972245773362196,"endowment":2.1972245773362196,"self_citation_contribution":0.32958368660043297,"citation_network_contribution":0.0,"self_endowment_contribution":0.32958368660043297,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":8,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9584,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":559162,"name":"Andrew Woodman","orcid":"0000-0001-7757-5492","position":1,"is_corresponding":false},{"id":559163,"name":"Kuo-Ming Lee","orcid":"0000-0002-3215-8342","position":2,"is_corresponding":false},{"id":463164,"name":"Ibrahim M. Moustafa","orcid":"0000-0002-5122-7569","position":3,"is_corresponding":false},{"id":559164,"name":"Fiona Fitzgerald","orcid":"0000-0001-7677-5118","position":4,"is_corresponding":false},{"id":559165,"name":"Peng-Nien Huang","orcid":"0000-0002-7345-2361","position":5,"is_corresponding":false},{"id":559166,"name":"Louis Kuijpers","orcid":"0000-0001-7245-3031","position":6,"is_corresponding":false},{"id":559936,"name":"Angela L. Perkins","orcid":null,"position":7,"is_corresponding":false},{"id":458585,"name":"Daniel A. Harki","orcid":"0000-0001-5950-931X","position":8,"is_corresponding":false},{"id":337326,"name":"Jamie J. Arnold","orcid":"0000-0002-2345-9776","position":9,"is_corresponding":false},{"id":559937,"name":"Belén Solano","orcid":null,"position":10,"is_corresponding":false},{"id":559167,"name":"Shin‐Ru Shih","orcid":"0000-0003-4874-401X","position":11,"is_corresponding":false},{"id":555983,"name":"Craig E. Cameron","orcid":"0000-0002-7564-5642","position":12,"is_corresponding":false},{"id":559168,"name":"Nynke H. Dekker","orcid":"0000-0003-4029-0973","position":13,"is_corresponding":false},{"id":559161,"name":"Richard Janissen","orcid":"0000-0003-0901-3433","position":0,"is_corresponding":true}],"reference_count":71,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-18T23:14:38.147936Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}