3L29

Crystal Structure of Zaire Ebola VP35 interferon inhibitory domain K319A/R322A mutant

Summary for 3L29

• Entry DOI: 10.2210/pdb3l29/pdb
• Related: 3FKE 3L25 3L26 3L27 3L28 3L2A
• Descriptor: Polymerase cofactor VP35, CHLORIDE ION (3 entities in total)
• Functional Keywords: rna binding domain, interferon antiviral evasion, rna replication, rna binding protein, transcription, host cytoplasm, interferon antiviral system evasion, rna-binding, virion
• Biological source: Zaire ebolavirus (ZEBOV)
• Cellular location: Virion: Q05127
• Total number of polymer chains: 2
• Total formula weight: 28203.52

Authors

Leung, D.W.,Ramanan, P.,Borek, D.M.,Amarasinghe, G.K. (deposition date: 2009-12-14, release date: 2010-02-02, Last modification date: 2023-09-06)

Primary citation

Prins, K.C.,Delpeut, S.,Leung, D.W.,Reynard, O.,Volchkova, V.A.,Reid, S.P.,Ramanan, P.,Cardenas, W.B.,Amarasinghe, G.K.,Volchkov, V.E.,Basler, C.F.
Mutations abrogating VP35 interaction with double-stranded RNA render ebola virus avirulent in guinea pigs.
J.Virol., 84:3004-3015, 2010

PubMed 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 critical EBOV virulence factor.

PubMed: 20071589
DOI: 10.1128/JVI.02459-09

Experimental method

X-RAY DIFFRACTION (1.7 Å)