Development of RNA aptamers targeting Ebola virus VP35

Jennifer M. Binning, Tianjiao Wang, Priya Luthra, Reed S. Shabman, Dominika M. Borek, Gai Liu, Wei Xu, Daisy W. Leung, Christopher F. Basler, Gaya K. Amarasinghe

Research output: Contribution to journalArticle

42 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)8406-8419
Number of pages14
JournalBiochemistry
Volume52
Issue number47
DOIs
StatePublished - Nov 26 2013

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Ebolavirus
Nucleotide Aptamers
Viral Proteins
Viruses
Nucleoproteins
Proteins
Interferons
Immune Evasion
Carrier Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Binning, J. M., Wang, T., Luthra, P., Shabman, R. S., Borek, D. M., Liu, G., ... Amarasinghe, G. K. (2013). Development of RNA aptamers targeting Ebola virus VP35. Biochemistry, 52(47), 8406-8419. https://doi.org/10.1021/bi400704d

Development of RNA aptamers targeting Ebola virus VP35. / Binning, Jennifer M.; Wang, Tianjiao; Luthra, Priya; Shabman, Reed S.; Borek, Dominika M.; Liu, Gai; Xu, Wei; Leung, Daisy W.; Basler, Christopher F.; Amarasinghe, Gaya K.

In: Biochemistry, Vol. 52, No. 47, 26.11.2013, p. 8406-8419.

Research output: Contribution to journalArticle

Binning, JM, Wang, T, Luthra, P, Shabman, RS, Borek, DM, Liu, G, Xu, W, Leung, DW, Basler, CF & Amarasinghe, GK 2013, 'Development of RNA aptamers targeting Ebola virus VP35', Biochemistry, vol. 52, no. 47, pp. 8406-8419. https://doi.org/10.1021/bi400704d
Binning JM, Wang T, Luthra P, Shabman RS, Borek DM, Liu G et al. Development of RNA aptamers targeting Ebola virus VP35. Biochemistry. 2013 Nov 26;52(47):8406-8419. https://doi.org/10.1021/bi400704d
Binning, Jennifer M. ; Wang, Tianjiao ; Luthra, Priya ; Shabman, Reed S. ; Borek, Dominika M. ; Liu, Gai ; Xu, Wei ; Leung, Daisy W. ; Basler, Christopher F. ; Amarasinghe, Gaya K. / Development of RNA aptamers targeting Ebola virus VP35. In: Biochemistry. 2013 ; Vol. 52, No. 47. pp. 8406-8419.
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