{"doi":"10.3390/v14071440","title":"Dynamic Interactions of Post Cleaved NS2B Cofactor and NS3 Protease Identified by Integrative Structural Approaches","abstract":"<jats:p>Diseases caused by flaviviruses such as dengue virus (DENV) and West Nile Virus (WNV), are a serious threat to public health. The flavivirus single-stranded RNA genome is translated into a polyprotein which is cleaved into three structural proteins and seven non-structural proteins by the viral and cellular proteases. Non-structural (NS) protein 3 is a multifunctional protein that has N-terminal protease and C-terminal helicase domains. The NS3 protease requires co-factor NS2B for enzymatic activity and folding. Due to its essential role in viral replication, NS2B-NS3 protease is an attractive target for antiviral drugs. Despite the availability of crystal structures, dynamic interactions of the N- and C-termini of NS2B co-factor have been elusive due to their flexible fold. In this study, we employ integrative structural approaches combined with biochemical assays to elucidate the dynamic interactions of the flexible DENV4 NS2B and NS3 N- and C-termini. We captured the crystal structure of self-cleaved DENV4 NS2B47NS3 protease in post cleavage state. The intermediate conformation adopted in the reported structure can be targeted by allosteric inhibitors. Comparison of our new findings from DENV4 against previously studied ZIKV NS2B-NS3 proteins reveals differences in NS2B-NS3 function between the two viruses. No inhibition of protease activity was observed for unlinked DENV NS2B-NS3 in presence of the cleavage site while ZIKV NS2B-NS3 cleavage inhibits protease activity. Another difference is that binding of the NS2B C-terminus to DENV4 eNS2B47NS3Pro active site is mediated via interactions with P4-P6 residues while for ZIKV, the binding of NS2B C-terminus to active site is mediated by P1-P3 residues. The mapping of NS2B N- and C-termini with NS3 indicates that these intermolecular interactions occur mainly on the beta-barrel 2 of the NS3 protease domain. Our integrative approach enables a comprehensive understanding of the folding and dynamic interactions of DENV NS3 protease and its cofactor NS2B.</jats:p>","journal":"Viruses","year":2022,"id":25680,"datarank":0.6504881086897234,"base_score":3.044522437723423,"endowment":3.044522437723423,"self_citation_contribution":0.4566783656585135,"citation_network_contribution":0.1938097430312099,"self_endowment_contribution":0.4566783656585135,"citer_contribution":0.1938097430312099,"corpus_percentile":null,"corpus_rank":null,"citation_count":20,"citer_count":17,"citers_with_citation_signal":9,"citers_with_endowment":9,"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":150452,"name":"Zheng Ser","orcid":"0000-0003-4297-7525","position":1,"is_corresponding":false},{"id":150453,"name":"Bing Liang Alvin Chew","orcid":"0000-0002-2748-9366","position":2,"is_corresponding":false},{"id":108969,"name":"Xin Li","orcid":"0000-0002-2122-7461","position":3,"is_corresponding":false},{"id":11852,"name":"Lili Wang","orcid":"0000-0002-8457-9616","position":4,"is_corresponding":false},{"id":150454,"name":"Radoslaw M. Sobota","orcid":null,"position":5,"is_corresponding":false},{"id":150456,"name":"Dahai Luo","orcid":null,"position":6,"is_corresponding":false},{"id":150458,"name":"Wint Wint Phoo","orcid":null,"position":7,"is_corresponding":false},{"id":150451,"name":"Jun-Ping Quek","orcid":"0000-0002-3827-8580","position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"base_score":3.044522437723423,"endowment":3.044522437723423,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"35891424","pmcid":"PMC9323329","openalex_id":"https://openalex.org/W4283747628","authors":[],"funders":[{"funder_name":"National Research Foundation","grant_id":"NRF2016NRF-CRP001-063","title":null},{"funder_name":"Fundação para a Ciência e a Tecnologia, I.P.","grant_id":"PTDC/CCI-INF/6762/2020","title":"MS3: New foundations for micro-services and serverless systems"}],"total_grants":2,"fwci":3.2483,"citation_percentile":0.91780654,"influential_citations":0,"citation_trend":[{"year":2022,"count":1},{"year":2023,"count":4},{"year":2024,"count":6},{"year":2025,"count":5},{"year":2026,"count":4}],"oa_status":"gold","license":"cc-by","oa_locations":[{"url":"https://www.mdpi.com/1999-4915/14/7/1440/pdf?version=1656582917","host_type":"journal"},{"url":"https://www.mdpi.com/1999-4915/14/7/1440/pdf?version=1656582917","host_type":"GOLD"},{"url":"https://www.mdpi.com/1999-4915/14/7/1440/pdf?version=1656582917","host_type":"publisher"},{"url":"https://www.mdpi.com/1999-4915/14/7/1440/pdf","host_type":"publisher"},{"url":"https://doi.org/10.3390/v14071440","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/35891424","host_type":"repository"},{"url":"https://doaj.org/article/bd11e71a03af46e9a5ad3d20cc3b1204","host_type":"repository"},{"url":"https://hdl.handle.net/10356/163157","host_type":"repository"},{"url":"https://dx.doi.org/10.3390/v14071440","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/9323329","host_type":"repository"},{"url":"https://europepmc.org/articles/PMC9323329","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC9323329?pdf=render","host_type":"Europe_PMC"},{"url":"http://dx.doi.org/10.3390/v14071440","host_type":""}],"fields_of_study":["Mosquito-borne diseases and control","HIV Research and Treatment","Protein Structure and Dynamics","Medicine","Biology","0301 basic medicine","03 medical and health sciences","0303 health sciences","Catalytic Domain","Flavivirus","Humans","Peptide Hydrolases","RNA Helicases","Serine Endopeptidases","Viral Nonstructural Proteins","Zika Virus","Zika Virus Infection"],"mesh_terms":["Zika Virus Infection","Zika Virus","Flavivirus","Humans","Peptide Hydrolases","Serine Endopeptidases","Viral Nonstructural Proteins","Catalytic Domain","RNA Helicases"],"keywords":["NS3","Protease","NS2-3 protease","Allosteric regulation","Flavivirus","Proteases","Cleavage (geology)","Biology","Chemistry","Dengue virus","Virology","Active site","Biochemistry","Enzyme","Dengue fever","Virus","X-ray crystallography","Denv","Viral Protease","Ns2b-ns3 Protease","Cross-linking Mass Spectrometry","Science::Medicine","Viral Nonstructural Proteins","Microbiology","Article","Catalytic Domain","Humans","Zika Virus Infection","Serine Endopeptidases","Zika Virus","QR1-502","RNA Helicases","Peptide Hydrolases"],"sdg_mappings":[{"sdg_number":3,"sdg_label":"3. 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