{"doi":"10.1128/jvi.78.6.2738-2748.2004","title":"Effect of Interaction between Hepatitis C Virus NS5A and NS5B on Hepatitis C Virus RNA Replication with the Hepatitis C Virus Replicon","abstract":"<jats:title>ABSTRACT</jats:title>\n          <jats:p>\n            Hepatitis C virus (HCV) NS5A has been reported to be important for the establishment of replication by adaptive mutations or localization, although its role in viral replication remains unclear. It was previously reported that NS5A interacts with NS5B via two regions of NS5A in the isolate JK-1 and modulates the activity of NS5B RdRp (Y. Shirota et al., J. Biol. Chem.,\n            <jats:bold>277:</jats:bold>\n            11149-11155, 2002), but the biological significance of this interaction has not been determined. In this study, we addressed the effect of this interaction on HCV RNA replication with an HCV replicon system derived from the isolate M1LE (H. Kishine et al., Biochem. Biophys. Res. Commun.,\n            <jats:bold>293:</jats:bold>\n            993-999, 2002). We constructed three internal deletion mutants, M1LE/5Adel-1 and M1LE/5Adel-2, each encoding NS5A which cannot bind NS5B, and M1LE/5Adel-3, encoding NS5A that can bind NS5B. After transfection into Huh-7 cells, M1LE/5Adel-3 was replication competent, but both M1LE/5Adel-1 and M1LE/5Adel-2 were not. Next we prepared 20 alanine-substituted clustered mutants within both NS5B-binding regions and examined the effect of these mutants on HCV RNA replication. Only 5 of the 20 mutants were replication competent. Subsequently, we introduced a point mutation, S225P, a deletion of S229, or S232I into NS5A and prepared cured Huh-7 cells that were cured of RNA replication by alpha interferon. Finally, with these point mutations and cured cells, we established a highly improved replicon system. In this system, only the same five mutants were replication competent. These results strongly suggest that the interaction between NS5A and NS5B is critical for HCV RNA replication in the HCV replicon system.\n          </jats:p>","journal":"Journal of Virology","year":2004,"id":25723,"datarank":4.790339870794133,"base_score":4.624972813284271,"endowment":4.624972813284271,"self_citation_contribution":0.6937459219926407,"citation_network_contribution":4.096593948801493,"self_endowment_contribution":0.6937459219926407,"citer_contribution":4.096593948801493,"corpus_percentile":null,"corpus_rank":null,"citation_count":101,"citer_count":96,"citers_with_citation_signal":84,"citers_with_endowment":84,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":149847,"name":"Makoto Hijikata","orcid":null,"position":1,"is_corresponding":false},{"id":135065,"name":"Hong Luo","orcid":null,"position":2,"is_corresponding":false},{"id":150611,"name":"Yuan Yuan Ma","orcid":null,"position":3,"is_corresponding":false},{"id":150612,"name":"Shuichi Kaneko","orcid":null,"position":4,"is_corresponding":false},{"id":133677,"name":"Kunitada Shimotohno","orcid":null,"position":5,"is_corresponding":false},{"id":135069,"name":"Seishi Murakami","orcid":null,"position":6,"is_corresponding":false},{"id":150610,"name":"Tetsuro Shimakami","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"base_score":4.624972813284271,"endowment":4.624972813284271,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"14990694","pmcid":"PMC353754","openalex_id":"https://openalex.org/W2139172730","authors":[],"funders":[],"total_grants":0,"fwci":4.9789,"citation_percentile":0.95741257,"influential_citations":5,"citation_trend":[{"year":2012,"count":5},{"year":2013,"count":7},{"year":2014,"count":4},{"year":2015,"count":7},{"year":2016,"count":4},{"year":2017,"count":4},{"year":2018,"count":3},{"year":2019,"count":2},{"year":2020,"count":5},{"year":2021,"count":2},{"year":2022,"count":1},{"year":2023,"count":2},{"year":2026,"count":2}],"oa_status":"green","license":"cc-by-nc-nd","oa_locations":[{"url":"https://kanazawa-u.repo.nii.ac.jp/records/38727","host_type":"repository"},{"url":"https://europepmc.org/articles/pmc353754?pdf=render","host_type":"GREEN"},{"url":"https://kanazawa-u.repo.nii.ac.jp/records/38727","host_type":"repository"},{"url":"https://journals.asm.org/doi/pdf/10.1128/JVI.78.6.2738-2748.2004","host_type":"publisher"},{"url":"https://doi.org/10.1128/jvi.78.6.2738-2748.2004","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/14990694","host_type":"repository"},{"url":"http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.653.8210","host_type":""},{"url":"http://europepmc.org/pmc/articles/PMC353754","host_type":"repository"},{"url":"https://kanazawa-u.repo.nii.ac.jp/?action=repository_uri&item_id=38727","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/353754","host_type":"repository"}],"fields_of_study":["Hepatitis C virus research","Hepatitis B Virus Studies","HIV Research and Treatment","Medicine","Biology","Amino Acid Sequence","Amino Acid Substitution","Animals","COS Cells","Cell Line, Tumor","Gene Expression Regulation, Viral","Hepacivirus","Humans","Molecular Sequence Data","RNA, Viral","Replicon","Sequence Deletion","Viral Nonstructural Proteins","Virus Replication","RNA-Dependent RNA Polymerase"],"mesh_terms":["Amino Acid Sequence","Animals","Humans","Molecular Sequence Data","Replicon","RNA-Dependent RNA Polymerase","RNA, Viral","Virus Replication","Gene Expression Regulation, Viral","Hepacivirus","Viral Nonstructural Proteins","Sequence Deletion","COS Cells","Amino Acid Substitution","Cell Line, Tumor"],"keywords":["NS5B","NS5A","Replicon","Biology","Viral replication","Virology","Hepatitis C virus","RNA","Mutant","Point mutation","RNA-dependent RNA polymerase","NS2-3 protease","Molecular biology","Virus","Hepacivirus","Gene","Genetics","Plasmid"],"sdg_mappings":[{"sdg_number":0,"sdg_label":"Good health and well-being"}],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[{"name":"gen"}],"source":"live","citation_network_status":"fetched"},"created_at":"2026-06-08T07:27:37.902253Z","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":[]}