Fragment screening targeting Ebola virus nucleoprotein C-terminal domain identifies lead candidates.

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  • Additional Information
    • Source:
      Publisher: Elsevier Country of Publication: Netherlands NLM ID: 8109699 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1872-9096 (Electronic) Linking ISSN: 01663542 NLM ISO Abbreviation: Antiviral Res Subsets: MEDLINE
    • Publication Information:
      Publication: Amsterdam : Elsevier
      Original Publication: [Amsterdam ; New York : Elsevier/North-Holland Biomedical Press, c1981-
    • Subject Terms:
      Structure-Activity Relationship*; Ebolavirus/*chemistry ; Nucleoproteins/*chemistry; Drug Discovery ; Ebolavirus/genetics ; Gene Library ; HEK293 Cells ; High-Throughput Screening Assays ; Humans ; Nucleoproteins/genetics ; Virus Replication
    • Abstract:
      The Ebola Virus is a causative agent of viral hemorrhagic fever outbreaks and a potential global health risk. The outbreak in West Africa (2013-2016) led to 11,000+ deaths and 30,000+ Ebola infected individuals. The current outbreak in the Democratic Republic of Congo (DRC) with 3000+ confirmed cases and 2000+ deaths attributed to Ebola virus infections provides a reminder that innovative countermeasures are still needed. Ebola virus encodes 7 open reading frames (ORFs). Of these, the nucleocapsid protein (eNP) encoded by the first ORF plays many significant roles, including a role in viral RNA synthesis. Here we describe efforts to target the C-terminal domain of eNP (eNP-CTD) that contains highly conserved residues 641-739 as a pan-Ebola antiviral target. Interactions of eNP-CTD with Ebola Viral Protein 30 (eVP30) and Viral Protein 40 (eVP40) have been shown to be crucial for viral RNA synthesis, virion formation, and virion transport. We used nuclear magnetic response (NMR)-based methods to screened the eNP-CTD against a fragment library. Perturbations of 1D 1 H NMR spectra identified of 48 of the 439 compounds screened as potential eNP CTD interactors. Subsequent analysis of these compounds to measure chemical shift perturbations in 2D 1 H, 15 N NMR spectra of 15 N-labeled protein identified six with low millimolar affinities. All six perturbed an area consisting mainly of residues at or near the extreme C-terminus that we named "Site 1" while three other sites were perturbed by other compounds. Our findings here demonstrate the potential utility of eNP as a target, several fragment hits, and provide an experimental pipeline to validate viral-viral interactions as potential panfiloviral inhibitor targets.
      Competing Interests: Declaration of competing interest None.
      (Copyright © 2020 The Author(s). Published by Elsevier B.V. All rights reserved.)
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    • Grant Information:
      T32 HL007575 United States HL NHLBI NIH HHS; R01 AI123926 United States AI NIAID NIH HHS; P01 AI120943 United States AI NIAID NIH HHS; P41 GM111135 United States GM NIGMS NIH HHS; P41 GM103399 United States GM NIGMS NIH HHS; R01 AI143292 United States AI NIAID NIH HHS
    • Contributed Indexing:
      Keywords: Drug development; Ebola virus; Fragment screening; Nucleoprotein C-terminal domain
    • Accession Number:
      0 (Nucleoproteins)
    • Publication Date:
      Date Created: 20200525 Date Completed: 20210716 Latest Revision: 20230123
    • Publication Date:
      20230123
    • Accession Number:
      PMC7894038
    • Accession Number:
      10.1016/j.antiviral.2020.104822
    • Accession Number:
      32446802