In Vitro Identification and In Vivo Confirmation of Inhibitors for Sweet Potato Chlorotic Stunt Virus RNA Silencing Suppressor, a Viral RNase III

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Wang , L , Pogue , S , Laamanen , K , Saarela , J , Poso , A , Laitinen , T & Valkonen , J P T 2021 , ' In Vitro Identification and In Vivo Confirmation of Inhibitors for Sweet Potato Chlorotic Stunt Virus RNA Silencing Suppressor, a Viral RNase III ' , Journal of Virology , vol. 95 , no. 12 , ARTN e00107-21 . https://doi.org/10.1128/JVI.00107-21

Title: In Vitro Identification and In Vivo Confirmation of Inhibitors for Sweet Potato Chlorotic Stunt Virus RNA Silencing Suppressor, a Viral RNase III
Author: Wang, Linping; Pogue, Sylvain; Laamanen, Karoliina; Saarela, Jani; Poso, Antti; Laitinen, Tuomo; Valkonen, Jari P. T.
Contributor organization: Department of Microbiology
Plant-Virus Interactions
Department of Agricultural Sciences
Institute for Molecular Medicine Finland
Biosciences
Viikki Plant Science Centre (ViPS)
Plant Pathology and Virology
Plant Production Sciences
Date: 2021-06
Language: eng
Number of pages: 15
Belongs to series: Journal of Virology
ISSN: 0022-538X
DOI: https://doi.org/10.1128/JVI.00107-21
URI: http://hdl.handle.net/10138/331648
Abstract: Sweet potato virus disease (SPVD), caused by synergistic infection of Sweet potato chlorotic stunt virus (SPCSV) and Sweet potato feathery mottle virus (SPFMV), is responsible for substantial yield losses all over the world. However, there are currently no approved treatments for this severe disease. The crucial role played by RNase III of SPCSV (CSR3) as an RNA silencing suppressor during the viruses' synergistic interaction in sweetpotato makes it an ideal drug target for developing antiviral treatment. In this study, high-throughput screening (HTS) of small molecular libraries targeting CSR3 was initiated by a virtual screen using Glide docking, allowing the selection of 6,400 compounds out of 136,353. We subsequently developed and carried out kinetic-based HTS using fluorescence resonance energy transfer technology, which isolated 112 compounds. These compounds were validated with dose-response assays including kinetic-based HTS and binding affinity assays using surface plasmon resonance and microscale thermophoresis. Finally, the interference of the selected compounds with viral accumulation was verified in planta. In summary, we identified five compounds belonging to two structural classes that inhibited CSR3 activity and reduced viral accumulation in plants. These results provide the foundation for developing antiviral agents targeting CSR3 to provide new strategies for controlling sweetpotato virus diseases. IMPORTANCE We report here a high-throughput inhibitor identification method that targets a severe sweetpotato virus disease caused by coinfection with two viruses (SPCSV and SPFMV). The disease is responsible for up to 90% yield losses. Specifically, we targeted the RNase III enzyme encoded by SPCSV, which plays an important role in suppressing the RNA silencing defense system of sweetpotato plants. Based on virtual screening, laboratory assays, and confirmation in planta, we identified five compounds that could be used to develop antiviral drugs to combat the most severe sweetpotato virus disease.
Subject: viral RNase III
RNA silencing suppressor
inhibitor identification
high-throughput screening
FRET
synergism
sweetpotato
SPCSV
EXOGENOUS APPLICATION
RIBONUCLEASE-III
MECHANISM
MOLECULES
BINDING
DSRNA
INTERFERENCE
RESISTANCE
ELIMINATION
DISCOVERY
1182 Biochemistry, cell and molecular biology
3121 General medicine, internal medicine and other clinical medicine
Peer reviewed: Yes
Rights: cc_by
Usage restriction: openAccess
Self-archived version: publishedVersion


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