1 research outputs found
Development of RNA Aptamers Targeting Ebola Virus VP35
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