skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Mutations Abrogating VP35 Interaction with Double-Stranded RNA Render Ebola Virus Avirulent in Guinea Pigs

Abstract

Ebola virus (EBOV) protein VP35 is a double-stranded RNA (dsRNA) binding inhibitor of host interferon (IFN)-{alpha}/{beta} responses that also functions as a viral polymerase cofactor. Recent structural studies identified key features, including a central basic patch, required for VP35 dsRNA binding activity. To address the functional significance of these VP35 structural features for EBOV replication and pathogenesis, two point mutations, K319A/R322A, that abrogate VP35 dsRNA binding activity and severely impair its suppression of IFN-{alpha}/{beta} production were identified. Solution nuclear magnetic resonance (NMR) spectroscopy and X-ray crystallography reveal minimal structural perturbations in the K319A/R322A VP35 double mutant and suggest that loss of basic charge leads to altered function. Recombinant EBOVs encoding the mutant VP35 exhibit, relative to wild-type VP35 viruses, minimal growth attenuation in IFN-defective Vero cells but severe impairment in IFN-competent cells. In guinea pigs, the VP35 mutant virus revealed a complete loss of virulence. Strikingly, the VP35 mutant virus effectively immunized animals against subsequent wild-type EBOV challenge. These in vivo studies, using recombinant EBOV viruses, combined with the accompanying biochemical and structural analyses directly correlate VP35 dsRNA binding and IFN inhibition functions with viral pathogenesis. Moreover, these studies provide a framework for the development of antivirals targeting this criticalmore » EBOV virulence factor.« less

Authors:
; ; ; ; ; ; ; ; ; ;  [1];  [2];  [2];  [2]
  1. (CNRS-INSERM)
  2. (
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
USDOE
OSTI Identifier:
1002793
Resource Type:
Journal Article
Resource Relation:
Journal Name: J. Virol.; Journal Volume: 84; Journal Issue: 03, 2010
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; 99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE; ANIMALS; ATTENUATION; CRYSTALLOGRAPHY; FUNCTIONALS; GENE MUTATIONS; GUINEA PIGS; IN VIVO; INTERFERON; MUTANTS; MUTATIONS; NUCLEAR MAGNETIC RESONANCE; PATHOGENESIS; POLYMERASES; PRODUCTION; PROTEINS; RNA; SPECTROSCOPY; VIRULENCE; VIRUSES

Citation Formats

Prins, Kathleen C., Delpeut, Sebastien, Leung, Daisy W., Reynard, Olivier, Volchkova, Valentina A., Reid, St. Patrick, Ramanan, Parameshwaran, Cárdenas, Washington B., Amarasinghe, Gaya K., Volchkov, Viktor E., Basler, Christopher F., Mount Sinai Hospital), LB-Ecuador), and Iowa State). Mutations Abrogating VP35 Interaction with Double-Stranded RNA Render Ebola Virus Avirulent in Guinea Pigs. United States: N. p., 2010. Web. doi:10.1128/JVI.02459-09.
Prins, Kathleen C., Delpeut, Sebastien, Leung, Daisy W., Reynard, Olivier, Volchkova, Valentina A., Reid, St. Patrick, Ramanan, Parameshwaran, Cárdenas, Washington B., Amarasinghe, Gaya K., Volchkov, Viktor E., Basler, Christopher F., Mount Sinai Hospital), LB-Ecuador), & Iowa State). Mutations Abrogating VP35 Interaction with Double-Stranded RNA Render Ebola Virus Avirulent in Guinea Pigs. United States. doi:10.1128/JVI.02459-09.
Prins, Kathleen C., Delpeut, Sebastien, Leung, Daisy W., Reynard, Olivier, Volchkova, Valentina A., Reid, St. Patrick, Ramanan, Parameshwaran, Cárdenas, Washington B., Amarasinghe, Gaya K., Volchkov, Viktor E., Basler, Christopher F., Mount Sinai Hospital), LB-Ecuador), and Iowa State). Mon . "Mutations Abrogating VP35 Interaction with Double-Stranded RNA Render Ebola Virus Avirulent in Guinea Pigs". United States. doi:10.1128/JVI.02459-09.
@article{osti_1002793,
title = {Mutations Abrogating VP35 Interaction with Double-Stranded RNA Render Ebola Virus Avirulent in Guinea Pigs},
author = {Prins, Kathleen C. and Delpeut, Sebastien and Leung, Daisy W. and Reynard, Olivier and Volchkova, Valentina A. and Reid, St. Patrick and Ramanan, Parameshwaran and Cárdenas, Washington B. and Amarasinghe, Gaya K. and Volchkov, Viktor E. and Basler, Christopher F. and Mount Sinai Hospital) and LB-Ecuador) and Iowa State)},
abstractNote = {Ebola virus (EBOV) protein VP35 is a double-stranded RNA (dsRNA) binding inhibitor of host interferon (IFN)-{alpha}/{beta} responses that also functions as a viral polymerase cofactor. Recent structural studies identified key features, including a central basic patch, required for VP35 dsRNA binding activity. To address the functional significance of these VP35 structural features for EBOV replication and pathogenesis, two point mutations, K319A/R322A, that abrogate VP35 dsRNA binding activity and severely impair its suppression of IFN-{alpha}/{beta} production were identified. Solution nuclear magnetic resonance (NMR) spectroscopy and X-ray crystallography reveal minimal structural perturbations in the K319A/R322A VP35 double mutant and suggest that loss of basic charge leads to altered function. Recombinant EBOVs encoding the mutant VP35 exhibit, relative to wild-type VP35 viruses, minimal growth attenuation in IFN-defective Vero cells but severe impairment in IFN-competent cells. In guinea pigs, the VP35 mutant virus revealed a complete loss of virulence. Strikingly, the VP35 mutant virus effectively immunized animals against subsequent wild-type EBOV challenge. These in vivo studies, using recombinant EBOV viruses, combined with the accompanying biochemical and structural analyses directly correlate VP35 dsRNA binding and IFN inhibition functions with viral pathogenesis. Moreover, these studies provide a framework for the development of antivirals targeting this critical EBOV virulence factor.},
doi = {10.1128/JVI.02459-09},
journal = {J. Virol.},
number = 03, 2010,
volume = 84,
place = {United States},
year = {Mon Oct 11 00:00:00 EDT 2010},
month = {Mon Oct 11 00:00:00 EDT 2010}
}