Discovery of novel inhibitors of RNA silencing suppressor P19 based on virtual screening

Hu, Fan; Lei, Rong; Deng, Yilun; Wang, Jun; Li, Guifen; Wang, Chao-Nan; Li, Zhihong; Zhu, Shuifang

doi:10.1039/c8ra01311j

Cited by 3 publications

(2 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Interestingly, this screening led to the identification of efficient antiviral agents against turnip mosaic virus (TuMV) later on (Fujiwara et al , 2013). Inhibitor screening for the RSS was mainly carried out by methods such as surface plasmon resonance and electrophoretic mobility shift assay (Sagan et al , 2007; Danielson et al , 2015; Hu et al , 2018).…”

Section: Introductionmentioning

confidence: 99%

Development of FRET‐based high‐throughput screening for viral RNase III inhibitors

Wang

Saarela

Poque

et al. 2020

Molecular Plant Pathology

View full text Add to dashboard Cite

The class 1 ribonuclease III (RNase III) encoded by Sweet potato chlorotic stunt virus (CSR3) suppresses RNA silencing in plant cells and thereby counters the host antiviral response by cleaving host small interfering RNAs, which are indispensable components of the plant RNA interference (RNAi) pathway. The synergy between sweet potato chlorotic stunt virus and sweet potato feathery mottle virus can reduce crop yields by 90%. Inhibitors of CSR3 might prove efficacious to counter this viral threat, yet no screen has been carried out to identify such inhibitors. Here, we report a novel high-throughput screening (HTS) assay based on fluorescence resonance energy transfer (FRET) for identifying inhibitors of CSR3. For monitoring CSR3 activity via HTS, we used a small interfering RNA substrate that was labelled with a FRETcompatible dye. The optimized HTS assay yielded 109 potential inhibitors of CSR3 out of 6,620 compounds tested from different small-molecule libraries. The three best inhibitor candidates were validated with a dose-response assay. In addition, a parallel screen of the selected candidates was carried out for a similar class 1 RNase III enzyme from Escherichia coli (EcR3), and this screen yielded a different set of inhibitors. Thus, our results show that the CSR3 and EcR3 enzymes were inhibited by distinct types of molecules, indicating that this HTS assay could be widely applied in drug discovery of class 1 RNase III enzymes. K E Y W O R D Sfluorescence resonance energy transfer, high-throughput screening, RNA silencing suppressor, RNase III, small interfering RNA

show abstract

Section: Introductionmentioning

confidence: 99%

Development of FRET‐based high‐throughput screening for viral RNase III inhibitors

Wang

Saarela

Poque

et al. 2020

Molecular Plant Pathology

View full text Add to dashboard Cite

show abstract

“…RNA silencing suppressors of Tombusvirus P19 (binding to siRNAs and therefore preventing their incorporation into RNA-induced silencing complex) have been targeted in inhibitors studies using an electrophoretic mobility shift assay, SPR, and/or fluorescence detection on Ni 2+ -NTA plates. These studies have led to the identification of chemical inhibitors interfering with its binding activity ( 40 – 42 ).…”

Section: Discussionmentioning

confidence: 99%

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

Wang

Poque

Laamanen

et al. 2021

J Virol

View full text Add to dashboard Cite

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 loss 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 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 a kinetic-based HTS using fluorescence resonance energy transfer technology that isolated 112 compounds. These compounds were validated with dose-response assays including the 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. Significance statement We report here a high-throughput inhibitor identification that targets a severe sweetpotato virus disease caused by co-infection with two viruses (SPCSV and SPFMV). The disease is responsible for up to 90% yield loss. 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.

show abstract

Small RNA and Degradome Deep Sequencing Reveal Regulatory Roles of MicroRNAs in Response to Sugarcane Mosaic Virus Infection on Two Contrasting Sugarcane Cultivars

Yuan,

Huang,

Pan

et al. 2024

MPMI

View full text Add to dashboard Cite

MicroRNAs (miRNAs) play an essential regulatory role in plant–virus interaction. However, few studies have focused on the roles of miRNAs and their targets after Sugarcane mosaic virus (SCMV) infection in sugarcane. To address this issue, we conducted small RNA and degradome sequencing on two contrasting sugarcanes (SCMV-resistant FG1 and susceptible Badila) infected by SCMV at five-time points. A total of 1578 miRNAs were profiled from 30 small RNA libraries, comprising 660 known miRNAs and 380 novel miRNAs. Differential expression analysis of miRNAs revealed that most were highly expressed during the SCMV exponential phase in Badila at 18h post-infection, with expression profiles positively correlated with virus replication dynamics, as observed through clustering. Analysis of degradome data indicated a higher number of differential miRNA targets in Badila compared to FG1 at 18 hours post-infection. Gene ontology (GO) enrichment analysis significantly enriched the stimulus-response pathway, suggesting negative regulatory roles to SCMV resistance. Specifically, miR160 exhibited upregulated expression patterns and validated in Badila through quantitative real-time PCR in the early stages of SCMV multiplication. Our research provides new insights into the dynamic response of plant miRNA and virus replication and contributes valuable information on the intricate interplay between miRNAs and SCMV infection dynamics.

show abstract

Discovery of novel inhibitors of RNA silencing suppressor P19 based on virtual screening

Cited by 3 publications

References 40 publications

Development of FRET‐based high‐throughput screening for viral RNase III inhibitors

Development of FRET‐based high‐throughput screening for viral RNase III inhibitors

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

Small RNA and Degradome Deep Sequencing Reveal Regulatory Roles of MicroRNAs in Response to Sugarcane Mosaic Virus Infection on Two Contrasting Sugarcane Cultivars

Contact Info

Product

Resources

About