A Sensitive in Vitro High-Throughput Screen to Identify Pan-filoviral Replication Inhibitors Targeting the VP35-NP Interface

Gai Liu, Peter J. Nash, Britney Johnson, Colette Pietzsch, Ma Xenia G. Ilagan, Alexander Bukreyev, Christopher F. Basler, Terry L. Bowlin, Donald T. Moir, Daisy W. Leung, Gaya K. Amarasinghe

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

The 2014 Ebola outbreak in West Africa, the largest outbreak on record, highlighted the need for novel approaches to therapeutics targeting Ebola virus (EBOV). Within the EBOV replication complex, the interaction between polymerase cofactor, viral protein 35 (VP35), and nucleoprotein (NP) is critical for viral RNA synthesis. We recently identified a peptide at the N-terminus of VP35 (termed NPBP) that is sufficient for interaction with NP and suppresses EBOV replication, suggesting that the NPBP binding pocket can serve as a potential drug target. Here we describe the development and validation of a sensitive high-throughput screen (HTS) using a fluorescence polarization assay. Initial hits from this HTS include the FDA-approved compound tolcapone, whose potency against EBOV infection was validated in a nonfluorescent secondary assay. High conservation of the NP-VP35 interface among filoviruses suggests that this assay has the capacity to identify pan-filoviral inhibitors for development as antivirals.

Original languageEnglish
Pages (from-to)190-198
Number of pages9
JournalACS Infectious Diseases
Volume3
Issue number3
DOIs
StatePublished - Mar 10 2017

Keywords

  • Ebola virus
  • VP35
  • fluorescence polarization assay
  • high-throughput screening
  • nucleoprotein

Fingerprint Dive into the research topics of 'A Sensitive in Vitro High-Throughput Screen to Identify Pan-filoviral Replication Inhibitors Targeting the VP35-NP Interface'. Together they form a unique fingerprint.

  • Cite this