A Sensitive in Vitro High-Throughput Screen To Identify Pan-filoviral Replication Inhibitors Targeting the VP35–NP Interface
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. Here, 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.
- Authors:
-
- Washington Univ. School of Medicine, St. Louis, MO (United States); Microbiotix Inc., Worcester, MA (United States)
- Microbiotix Inc., Worcester, MA (United States)
- Washington Univ. School of Medicine, St. Louis, MO (United States)
- Univ. of Texas Medical Branch, Galveston, TX (United States)
- Georgia State Univ., Atlanta, GA (United States)
- Publication Date:
- Research Org.:
- Argonne National Laboratory (ANL), Argonne, IL (United States)
- Sponsoring Org.:
- National Institutes of Health (NIH)
- OSTI Identifier:
- 1351401
- Grant/Contract Number:
- R01AI123926; R01 AI114654; U19AI109945; P01AI120943
- Resource Type:
- Journal Article: Accepted Manuscript
- Journal Name:
- ACS Infectious Diseases
- Additional Journal Information:
- Journal Volume: 3; Journal Issue: 3; Journal ID: ISSN 2373-8227
- Publisher:
- American Chemical Society (ACS)
- Country of Publication:
- United States
- Language:
- ENGLISH
- Subject:
- 60 APPLIED LIFE SCIENCES; Ebola virus; nucleoprotein; VP35; fluorescence polarization assay; high-throughput screening; DNA replication; peptides and proteins; fluorescence; assays; inhibitors
Citation Formats
Liu, Gai, Nash, Peter J., Johnson, Britney, Pietzsch, Colette, Ilagan, Ma. Xenia G., Bukreyev, Alexander, Basler, Christopher F., Bowlin, Terry L., Moir, Donald T., Leung, Daisy W., and Amarasinghe, Gaya K. A Sensitive in Vitro High-Throughput Screen To Identify Pan-filoviral Replication Inhibitors Targeting the VP35–NP Interface. United States: N. p., 2017.
Web. doi:10.1021/acsinfecdis.6b00209.
Liu, Gai, Nash, Peter J., Johnson, Britney, Pietzsch, Colette, Ilagan, Ma. Xenia G., Bukreyev, Alexander, Basler, Christopher F., Bowlin, Terry L., Moir, Donald T., Leung, Daisy W., & Amarasinghe, Gaya K. A Sensitive in Vitro High-Throughput Screen To Identify Pan-filoviral Replication Inhibitors Targeting the VP35–NP Interface. United States. https://doi.org/10.1021/acsinfecdis.6b00209
Liu, Gai, Nash, Peter J., Johnson, Britney, Pietzsch, Colette, Ilagan, Ma. Xenia G., Bukreyev, Alexander, Basler, Christopher F., Bowlin, Terry L., Moir, Donald T., Leung, Daisy W., and Amarasinghe, Gaya K. 2017.
"A Sensitive in Vitro High-Throughput Screen To Identify Pan-filoviral Replication Inhibitors Targeting the VP35–NP Interface". United States. https://doi.org/10.1021/acsinfecdis.6b00209. https://www.osti.gov/servlets/purl/1351401.
@article{osti_1351401,
title = {A Sensitive in Vitro High-Throughput Screen To Identify Pan-filoviral Replication Inhibitors Targeting the VP35–NP Interface},
author = {Liu, Gai and Nash, Peter J. and Johnson, Britney and Pietzsch, Colette and Ilagan, Ma. Xenia G. and Bukreyev, Alexander and Basler, Christopher F. and Bowlin, Terry L. and Moir, Donald T. and Leung, Daisy W. and Amarasinghe, Gaya K.},
abstractNote = {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. Here, 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.},
doi = {10.1021/acsinfecdis.6b00209},
url = {https://www.osti.gov/biblio/1351401},
journal = {ACS Infectious Diseases},
issn = {2373-8227},
number = 3,
volume = 3,
place = {United States},
year = {Thu Feb 02 00:00:00 EST 2017},
month = {Thu Feb 02 00:00:00 EST 2017}
}
Web of Science
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