Document Detail


Development of RNA aptamers targeting Ebola virus VP35.
MedLine Citation:
PMID:  24067086     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
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
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2013-11-14
Journal Detail:
Title:  Biochemistry     Volume:  52     ISSN:  1520-4995     ISO Abbreviation:  Biochemistry     Publication Date:  2013 Nov 
Date Detail:
Created Date:  2013-11-26     Completed Date:  2014-01-30     Revised Date:  2014-02-03    
Medline Journal Info:
Nlm Unique ID:  0370623     Medline TA:  Biochemistry     Country:  United States    
Other Details:
Languages:  eng     Pagination:  8406-19     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Antiviral Agents / chemistry*,  metabolism,  pharmacology
Aptamers, Nucleotide / chemistry*,  metabolism,  pharmacology
Binding, Competitive
Conserved Sequence
Ebolavirus / drug effects,  metabolism
Hemorrhagic Fever, Ebola / drug therapy,  metabolism
Kinetics
Molecular Targeted Therapy
Mutant Proteins / antagonists & inhibitors,  chemistry,  metabolism
Nucleic Acid Conformation
Nucleoproteins / metabolism
Protein Conformation
Protein Interaction Domains and Motifs
RNA / chemistry*,  metabolism,  pharmacology
RNA, Double-Stranded / metabolism
Recombinant Proteins / chemistry,  metabolism
SELEX Aptamer Technique
Species Specificity
Viral Regulatory and Accessory Proteins / antagonists & inhibitors*,  chemistry,  genetics,  metabolism
Grant Support
ID/Acronym/Agency:
R01 AI059536/AI/NIAID NIH HHS; R01 AI081914/AI/NIAID NIH HHS; R01 GM053163/GM/NIGMS NIH HHS; R01AI059536/AI/NIAID NIH HHS; R01AI081914/AI/NIAID NIH HHS; U54 AI057160/AI/NIAID NIH HHS; U54AI057160/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/Antiviral Agents; 0/Aptamers, Nucleotide; 0/Mutant Proteins; 0/Nucleoproteins; 0/RNA, Double-Stranded; 0/Recombinant Proteins; 0/VP35 protein, filovirus; 0/Viral Regulatory and Accessory Proteins; 63231-63-0/RNA

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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