4IJF
Crystal structure of the Zaire ebolavirus VP35 interferon inhibitory domain K222A/R225A/K248A/K251A mutant
Summary for 4IJF
| Entry DOI | 10.2210/pdb4ijf/pdb |
| Related | 3FKE 4IJE |
| Descriptor | Polymerase cofactor VP35 (2 entities in total) |
| Functional Keywords | ifn inhibition, polymerase cofactor, rna binding protein, interferon antagonism, dsrna, virus, viral protein |
| Biological source | Zaire ebolavirus (ZEBOV) |
| Cellular location | Virion: Q05127 |
| Total number of polymer chains | 1 |
| Total formula weight | 13852.99 |
| Authors | Binning, J.B.,Wang, T.,Leung, D.W.,Xu, W.,Borek, D.,Amarasinghe, G.K. (deposition date: 2012-12-21, release date: 2013-10-09, Last modification date: 2024-10-09) |
| Primary citation | Binning, J.M.,Wang, T.,Luthra, P.,Shabman, R.S.,Borek, D.M.,Liu, G.,Xu, W.,Leung, D.W.,Basler, C.F.,Amarasinghe, G.K. Development of RNA Aptamers Targeting Ebola Virus VP35. Biochemistry, 52:8406-8419, 2013 Cited by PubMed Abstract: Viral protein 35 (VP35), encoded by filoviruses, is a multifunctional dsRNA binding protein that plays important roles in viral replication, innate immune evasion, and pathogenesis. The multifunctional nature of these proteins also presents opportunities to develop countermeasures that target distinct functional regions. However, functional validation and the establishment of therapeutic approaches toward such multifunctional proteins, particularly for nonenzymatic targets, are often challenging. Our previous work on filoviral VP35 proteins defined conserved basic residues located within its C-terminal dsRNA binding interferon (IFN) inhibitory domain (IID) as important for VP35 mediated IFN antagonism and viral polymerase cofactor functions. In the current study, we used a combination of structural and functional data to determine regions of Ebola virus (EBOV) VP35 (eVP35) to target for aptamer selection using SELEX. Select aptamers, representing, two distinct classes, were further characterized based on their interaction properties to eVP35 IID. These results revealed that these aptamers bind to distinct regions of eVP35 IID with high affinity (10-50 nM) and specificity. These aptamers can compete with dsRNA for binding to eVP35 and disrupt the eVP35-nucleoprotein (NP) interaction. Consistent with the ability to antagonize the eVP35-NP interaction, select aptamers can inhibit the function of the EBOV polymerase complex reconstituted by the expression of select viral proteins. Taken together, our results support the identification of two aptamers that bind filoviral VP35 proteins with high affinity and specificity and have the capacity to potentially function as filoviral VP35 protein inhibitors. PubMed: 24067086DOI: 10.1021/bi400704d PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.506 Å) |
Structure validation
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