Simple jQuery Dropdowns
Please use this identifier to cite or link to this item:
Title: Interference of dengue replication by blocking the access of 3′ SL RNA to the viral RNA-dependent RNA polymerase
Authors: Chairat Tunghirun
Veerakorn Narkthong
Wanpen Chaicumpa
Sarin Chimnaronk
Mahidol University
Faculty of Medicine, Siriraj Hospital, Mahidol University
Keywords: Immunology and Microbiology
Issue Date: 1-Oct-2020
Citation: Antiviral Research. Vol.182, (2020)
Abstract: © 2020 Elsevier B.V. The four circulating serotypes of dengue virus (DENV) occasionally cause potentially fetal symptoms of severe dengue, which there is currently no specific treatment available. Extensive efforts have been made to inhibit viral replication processes by impeding the activity of an exclusive RNA-dependent RNA polymerase (RdRp) in the viral non-structural protein 5 (NS5). In our earlier work, we identified the characteristic, specific interaction between the C-terminal thumb subdomain of RdRp and an apical loop in the 3′ stem-loop (SL) element in the DENV RNA genome, which is fundamental for viral replication. Here, we demonstrated a new approach for interfering viral replication via blocking of 3′ SL RNA binding to RdRp by the single-chain variable fragments (scFvs). We isolated and cloned 3 different human scFvs that bound to RdRp from DENV serotype 2 and interfered with 3′ SL-binding, utilizing a combination of phage-display panning and Alpha methods. When tagged with a cell penetrating peptide, a selected scFv clone, 2E3, entered cells and partially colocalized with NS5 in the cytoplasm of infected HuH-7 cells. 2E3 significantly inhibited DENV RNA replication with sub-nanomolar EC50 values and significantly reduced the production of infectious particles. The molecular docking models suggested that 2E3 recognized both palm and thumb subdomains of RdRp, and interacted with Lys841, a key residue involved in RNA binding. Our results provide a new potential therapeutic molecule specific for flaviviral infection.
ISSN: 18729096
Appears in Collections:Scopus 2020

Files in This Item:
There are no files associated with this item.

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.