{"doi":"10.3390/v12010105","title":"Reporter Assays for Ebola Virus Nucleoprotein Oligomerization, Virion-Like Particle Budding, and Minigenome Activity Reveal the Importance of Nucleoprotein Amino Acid Position 111","abstract":"For highly pathogenic viruses, reporter assays that can be rapidly performed are critically needed to identify potentially functional mutations for further study under maximal containment (e.g., biosafety level 4 [BSL-4]). The Ebola virus nucleoprotein (NP) plays multiple essential roles during the viral life cycle, yet few tools exist to study the protein under BSL-2 or equivalent containment. Therefore, we adapted reporter assays to measure NP oligomerization and virion-like particle (VLP) production in live cells and further measured transcription and replication using established minigenome assays. As a proof-of-concept, we examined the NP-R111C substitution, which emerged during the 2013‒2016 Western African Ebola virus disease epidemic and rose to high frequency. NP-R111C slightly increased NP oligomerization and VLP budding but slightly decreased transcription and replication. By contrast, a synthetic charge-reversal mutant, NP-R111E, greatly increased oligomerization but abrogated transcription and replication. These results are intriguing in light of recent structures of NP oligomers, which reveal that the neighboring residue, K110, forms a salt bridge with E349 on adjacent NP molecules. By developing and utilizing multiple reporter assays, we find that the NP-111 position mediates a complex interplay between NP's roles in protein structure, virion budding, and transcription and replication.","journal":"Viruses","year":2020,"id":86354,"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":13,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9661,"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":284587,"name":"William E. Diehl","orcid":"0000-0001-7943-1214","position":1,"is_corresponding":false},{"id":440220,"name":"Yíngyún Caì","orcid":"0000-0002-1101-0920","position":2,"is_corresponding":false},{"id":218417,"name":"Courtney L. Finch","orcid":"0000-0002-3674-3282","position":3,"is_corresponding":false},{"id":440221,"name":"Chidiebere Akusobi","orcid":"0000-0002-1611-0015","position":4,"is_corresponding":false},{"id":279924,"name":"Robert N. Kirchdoerfer","orcid":null,"position":5,"is_corresponding":false},{"id":109939,"name":"Laura Bollinger","orcid":"0000-0002-6676-8894","position":6,"is_corresponding":false},{"id":62163,"name":"S. F. Schaffner","orcid":"0000-0001-6699-3568","position":7,"is_corresponding":false},{"id":440222,"name":"Elizabeth A. Brown","orcid":"0000-0002-0832-3758","position":8,"is_corresponding":false},{"id":104945,"name":"Erica Ollmann Saphire","orcid":"0000-0002-1206-7451","position":9,"is_corresponding":false},{"id":234927,"name":"Kristian G. Andersen","orcid":"0000-0001-6431-5982","position":10,"is_corresponding":false},{"id":109882,"name":"Jens H. Kuhn","orcid":"0000-0002-7800-6045","position":11,"is_corresponding":false},{"id":284590,"name":"Jeremy Luban","orcid":"0000-0001-5650-4054","position":12,"is_corresponding":false},{"id":7155,"name":"Pardis C. Sabeti","orcid":"0000-0002-9843-1890","position":13,"is_corresponding":false},{"id":109804,"name":"Aaron E. Lin","orcid":"0000-0001-7400-4125","position":0,"is_corresponding":true}],"reference_count":83,"raw_metadata":null,"created_at":"2026-07-18T21:58:41.997408Z","pmid":"31952352","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":[]}