Mutations abrogating VP35 interaction with double-stranded RNA render Ebola virus avirulent in guinea pigs

Kathleen C. Prins, Sebastien Delpeut, Daisy W. Leung, Olivier Reynard, Valentina A. Volchkova, St Patrick Reid, Parameshwaran Ramanan, Washington B. Cárdenas, Gaya K. Amarasinghe, Viktor E. Volchkov, Christopher F. Basler

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98 Citations (Scopus)

Abstract

Ebola virus (EBOV) protein VP35 is a double-stranded RNA (dsRNA) binding inhibitor of host interferon (IFN)-α/β 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;/β 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.

Original languageEnglish (US)
Pages (from-to)3004-3015
Number of pages12
JournalJournal of virology
Volume84
Issue number6
DOIs
StatePublished - Mar 1 2010

Fingerprint

Ebolavirus
Double-Stranded RNA
double-stranded RNA
guinea pigs
Interferons
interferons
Guinea Pigs
Viruses
mutation
Mutation
viruses
mutants
virulence
pathogenesis
Vero Cells
X Ray Crystallography
Virulence Factors
Virus Replication
Point Mutation
interferon-alpha

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Prins, K. C., Delpeut, S., Leung, D. W., Reynard, O., Volchkova, V. A., Reid, S. P., ... Basler, C. F. (2010). Mutations abrogating VP35 interaction with double-stranded RNA render Ebola virus avirulent in guinea pigs. Journal of virology, 84(6), 3004-3015. https://doi.org/10.1128/JVI.02459-09

Mutations abrogating VP35 interaction with double-stranded RNA render Ebola virus avirulent in guinea pigs. / 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.

In: Journal of virology, Vol. 84, No. 6, 01.03.2010, p. 3004-3015.

Research output: Contribution to journalArticle

Prins, KC, Delpeut, S, Leung, DW, Reynard, O, Volchkova, VA, Reid, SP, Ramanan, P, Cárdenas, WB, Amarasinghe, GK, Volchkov, VE & Basler, CF 2010, 'Mutations abrogating VP35 interaction with double-stranded RNA render Ebola virus avirulent in guinea pigs', Journal of virology, vol. 84, no. 6, pp. 3004-3015. https://doi.org/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. / Mutations abrogating VP35 interaction with double-stranded RNA render Ebola virus avirulent in guinea pigs. In: Journal of virology. 2010 ; Vol. 84, No. 6. pp. 3004-3015.
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