title: Structural basis for the identification of the N-terminal domain of coronavirus nucleocapsid protein as an antiviral target.
abstract: Coronaviruses (CoVs) cause numerous diseases, including Middle East respiratory syndrome and severe acute respiratory syndrome, generating significant health-related and economic consequences. CoVs encode the nucleocapsid (N) protein, a major structural protein that plays multiple roles in the virus replication cycle and forms a ribonucleoprotein complex with the viral RNA through the N protein's N-terminal domain (N-NTD). Using human CoV-OC43 (HCoV-OC43) as a model for CoV, we present the 3D structure of HCoV-OC43 N-NTD complexed with ribonucleoside 5'-monophosphates to identify a distinct ribonucleotide-binding pocket. By targeting this pocket, we identified and developed a new coronavirus N protein inhibitor, N-(6-oxo-5,6-dihydrophenanthridin-2-yl)(N,N-dimethylamino)acetamide hydrochloride (PJ34), using virtual screening; this inhibitor reduced the N protein's RNA-binding affinity and hindered viral replication. We also determined the crystal structure of the N-NTD-PJ34 complex. On the basis of these findings, we propose guidelines for developing new N protein-based antiviral agents that target CoVs.
authors.affiliation: College of Life Science, ‡Institute of Genomics and Bioinformatics, and §Agriculture Biotechnology Center, National Chung Hsing University , Taichung 40254, Taiwan.
authors.forename: Shing-Yen
authors.initials: SY
authors.lastname: Lin
authors.forename: Chia-Ling
authors.initials: CL
authors.lastname: Liu
authors.forename: Yu-Ming
authors.initials: YM
authors.lastname: Chang
authors.forename: Jincun
authors.initials: J
authors.lastname: Zhao
authors.forename: Stanley
authors.initials: S
authors.lastname: Perlman
authors.forename: Ming-Hon
authors.initials: MH
authors.lastname: Hou
mesh_terms: D000998:Antiviral Agents / Q000737:chemistry* / Q000494:pharmacology*; D017934:Coronavirus / Q000187:drug effects*; D000086462:Coronavirus Nucleocapsid Proteins; D015195:Drug Design; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D008968:Molecular Conformation; D016297:Mutagenesis, Site-Directed; D019590:Nucleocapsid Proteins / Q000737:chemistry* / Q000187:drug effects*; D010616:Phenanthrenes / Q000138:chemical synthesis* / Q000494:pharmacology*; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D012367:RNA, Viral / Q000187:drug effects / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D013329:Structure-Activity Relationship; D014779:Virus Replication / Q000187:drug effects; D014961:X-Ray Diffraction
chemical_list: D000998:Antiviral Agents; D000086462:Coronavirus Nucleocapsid Proteins; C434926:N-(oxo-5,6-dihydrophenanthridin-2-yl)-N,N-dimethylacetamide hydrochloride; D019590:Nucleocapsid Proteins; D010616:Phenanthrenes; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D012367:RNA, Viral