{"doi":"10.1101/2023.11.29.569222","title":"Structural and functional characterisation of the interaction between the influenza A virus RNA polymerase and the CTD of host RNA Polymerase II","abstract":"<jats:title>ABSTRACT</jats:title>\n                <jats:p>Influenza A viruses (IAV), causing seasonal epidemics and occasional pandemics, rely on interactions with host proteins for their RNA genome transcription and replication. The viral RNA polymerase utilizes host RNA polymerase II (Pol II) and interacts with the serine 5 phosphorylated (pS5) C-terminal domain (CTD) of Pol II to initiate transcription. Our study, using single-particle electron cryomicroscopy (cryo-EM), reveals the structure of the 1918 pandemic IAV polymerase bound to a synthetic pS5 CTD heptad repeat peptide. The structure shows that the CTD peptide binds at the C-terminal domain of the PA viral polymerase subunit (PA-C) and reveals a previously unobserved position of the 627 domain of the PB2 subunit near the CTD. We identify crucial residues of the CTD peptide mediating interactions with positively charged cavities on PA-C, explaining the preference of the viral polymerase for pS5 CTD. Functional analysis of mutants targeting the CTD-binding site within PA-C reveals reduced transcriptional function with normal replication, while other mutants display defects in both transcription and replication, highlighting the multifunctional role of PA-C in viral RNA synthesis. Our study provides insights into the structural and functional aspects of the influenza virus polymerase-host Pol II interaction and identifies a target for antiviral development.</jats:p>","journal":null,"year":null,"id":630753,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":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":1634247,"name":"Alaa Baazaoui","orcid":null,"position":1,"is_corresponding":false},{"id":1634249,"name":"Marek Šebesta","orcid":null,"position":2,"is_corresponding":false},{"id":1634250,"name":"Richard Štefl","orcid":null,"position":3,"is_corresponding":false},{"id":1634252,"name":"Loic Carrique","orcid":null,"position":4,"is_corresponding":false},{"id":181179,"name":"Ervin Fodor","orcid":null,"position":5,"is_corresponding":false},{"id":1634254,"name":"Jonathan Grimes","orcid":null,"position":6,"is_corresponding":false},{"id":1634246,"name":"Jeremy Keown","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Structural and functional characterisation of the interaction between the influenza A virus RNA polymerase and the CTD of host RNA Polymerase II","abstract":"<jats:title>ABSTRACT</jats:title>\n                <jats:p>Influenza A viruses (IAV), causing seasonal epidemics and occasional pandemics, rely on interactions with host proteins for their RNA genome transcription and replication. The viral RNA polymerase utilizes host RNA polymerase II (Pol II) and interacts with the serine 5 phosphorylated (pS5) C-terminal domain (CTD) of Pol II to initiate transcription. Our study, using single-particle electron cryomicroscopy (cryo-EM), reveals the structure of the 1918 pandemic IAV polymerase bound to a synthetic pS5 CTD heptad repeat peptide. The structure shows that the CTD peptide binds at the C-terminal domain of the PA viral polymerase subunit (PA-C) and reveals a previously unobserved position of the 627 domain of the PB2 subunit near the CTD. We identify crucial residues of the CTD peptide mediating interactions with positively charged cavities on PA-C, explaining the preference of the viral polymerase for pS5 CTD. Functional analysis of mutants targeting the CTD-binding site within PA-C reveals reduced transcriptional function with normal replication, while other mutants display defects in both transcription and replication, highlighting the multifunctional role of PA-C in viral RNA synthesis. Our study provides insights into the structural and functional aspects of the influenza virus polymerase-host Pol II interaction and identifies a target for antiviral development.</jats:p>","is_dataset_classified":null,"base_score":0.0,"endowment":0.0,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"19767382","pmcid":null,"openalex_id":"https://openalex.org/W4389153874","authors":[],"funders":[{"funder_name":"Wellcome Trust","grant_id":"203141/Z/16/Z","title":null},{"funder_name":"European Commission","grant_id":"200835","title":"ASSESSMENT OF HEARING IN THE ELDERLY: AGING AND DEGENERATION - INTEGRATION THROUGH IMMEDIATE INTERVENTION"},{"funder_name":"Wellcome Trust","grant_id":"222510","title":"Replication and Assembly of Influenza virus"},{"funder_name":"Wellcome Trust","grant_id":"203141","title":"Human Genetics and Disease Biology: Core Renewal for the Wellcome Trust Centre for Human Genetics"},{"funder_name":"UK Research and Innovation","grant_id":"MR/X008312/1","title":"Transcription, replication, trafficking and assembly of the influenza virus RNA genome"},{"funder_name":"UK Research and Innovation","grant_id":"BB/M011224/1","title":"The Oxford Interdisciplinary Bioscience Doctoral Training Partnership"},{"funder_name":"UK Research and Innovation","grant_id":"MR/R009945/1","title":"Structure-function relationships of the influenza virus RNA polymerase: influence on virulence, host restriction and innate immune responses"},{"funder_name":"Wellcome Trust","grant_id":"093305","title":"Purchase of electron microscopic infrastructure to establish a multi-user facility for cryo electron microscopy, in particular cryo electron tomography."}],"total_grants":8,"fwci":null,"citation_percentile":null,"influential_citations":0,"citation_trend":[],"oa_status":"green","license":"cc-by","oa_locations":[{"url":"https://www.biorxiv.org/content/biorxiv/early/2023/11/29/2023.11.29.569222.full.pdf","host_type":"repository"},{"url":"https://www.biorxiv.org/content/biorxiv/early/2023/11/29/2023.11.29.569222.full.pdf","host_type":"repository"},{"url":"https://syndication.highwire.org/content/doi/10.1101/2023.11.29.569222","host_type":"publisher"},{"url":"http://dx.doi.org/10.1101/2023.11.29.569222","host_type":"repository"},{"url":"https://europepmc.org/article/PPR/PPR766346","host_type":"Europe_PMC"},{"url":"https://europepmc.org/api/fulltextRepo?pprId=PPR766346&type=FILE&fileName=EMS191879-pdf.pdf&mimeType=application/pdf","host_type":"Europe_PMC"},{"url":"https://doi.org/10.1128/jvi.00138-24","host_type":""},{"url":"https://doi.org/10.1101/2023.11.29.569222","host_type":""},{"url":"https://pubmed.ncbi.nlm.nih.gov/38563748","host_type":""},{"url":"http://dx.doi.org/10.1128/jvi.00138-24","host_type":""},{"url":"https://ora.ox.ac.uk/objects/uuid:faf5bf6c-090d-4740-9c4a-3b9fbef7661c","host_type":""},{"url":"https://is.muni.cz/publication/2419757","host_type":""}],"fields_of_study":["Influenza Virus Research Studies","RNA and protein synthesis mechanisms","RNA Research and Splicing","0301 basic medicine","0303 health sciences","03 medical and health sciences"],"mesh_terms":[],"keywords":["RNA polymerase II","Polymerase","Biology","CTD","Transcription (linguistics)","RNA polymerase","Transcription factor II D","Influenza A virus","Protein subunit","Molecular biology","C-terminus","RNA-dependent RNA polymerase","RNA polymerase I","Virology","RNA","Cell biology","Virus","Genetics","Gene","Gene expression","Promoter","Models, Molecular","Transcription, Genetic","Cryoelectron Microscopy","Virus Replication","Genome Replication and Regulation of Viral Gene Expression","Viral Proteins","influenza; RNA polymerases; transcription; RNA polymerase II; CTD","Protein Domains","Influenza, Human","Humans","RNA, Viral","Phosphorylation","Protein Binding"],"sdg_mappings":[{"sdg_number":3,"sdg_label":"3. 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