Dataset Information

Proxitome profiling of a viral suppressors of RNA silencing identifies the DEAD-box RNA helicase RH20 as a positive regulator of antiviral RNA interference in Nicotiana benthamiana

ABSTRACT: In the prolonged arms race between viruses and their host plants, viruses usually encode viral suppressors of RNA silencing (VSR) to overcome the hosts' defense mechanism by inhibiting RNAi. Barley stripe mosaic virus (BSMV) is an RNA virus characterized by a positive single-stranded genetic structure, comprising three distinct genomic RNAs labeled as α, β, and γ. The multifunctional protein γb within BSMV’s RNAγ acts as a potent VSR, yet its underlying molecular mechanism requires further exploration. Previous studies on BSMV interacting proteins were conducted exclusively in yeast systems. We aim to identify the interacting proteins of γb under authentic conditions, specifically, the protein group interacting in the context of BSMV infection. The emerging proximity labeling (PL) technology offers several advantages, such as the identification of transiently interacting proteins, membrane proteins, and being non-toxic to plants. Consequently, this determines that PL technology may aid in resolving the challenge of identifying the interacting protein group of VSR in the context of BSMV infection.

ORGANISM(S): Nicotiana Benthamiana

SUBMITTER: Yongliang Zhang

PROVIDER: PXD049705 | iProX | Tue Feb 20 00:00:00 GMT 2024

REPOSITORIES: iProX

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Publications

DEAD-box RNA helicase RH20 positively regulates RNAi-based antiviral immunity in plants by associating with SGS3/RDR6 bodies.

Wen Zhiyan Z Hu Rujian R Pi Qinglin Q Zhang Dingliang D Duan Jiangning J Li Zhen Z Li Qian Q Zhao Xiaoyun X Yang Meng M Zhao Xiaofei X Liu Deshui D Su Zhen Z Li Dawei D Zhang Yongliang Y

Plant biotechnology journal 20240821 12

RNA silencing plays a crucial role in defending against viral infections in diverse eukaryotic hosts. Despite extensive studies on core components of the antiviral RNAi pathway such as DCLs, AGOs and RDRs proteins, host factors involved in antiviral RNAi remain incompletely understood. In this study, we employed the proximity labelling approach to identify the host factors required for antiviral RNAi in Nicotiana benthamiana. Using the barley stripe mosaic virus (BSMV)-encoded γb, a viral suppre ...[more]

PMID: 39166471

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Project description:RNA silencing is one of the main defense mechanisms employed by plants to fight viruses. In change, viruses have evolved silencing suppressor proteins to neutralize antiviral silencing. Since the endogenous and antiviral functions of RNA silencing pathway rely on common components, it was suggested that viral suppressors interfere with endogenous silencing pathway contributing to viral symptom development. In this work, we aimed to understand the effects of the tombusviral p19 suppressor on endogenous and antiviral silencing during genuine virus infection. We showed that ectopically expressed p19 sequesters endogenous small RNAs (sRNAs) in the absence, but not in the presence of virus infection. Our presented data question the generalized model in which the sequestration of endogenous sRNAs by the viral suppressor contributes to the viral symptom development. We further showed that p19 preferentially binds the perfectly-paired ds-viral small interfering RNAs (vsiRNAs) but does not select based on their sequence or the type of the 5’ nucleotide. Finally, co-immunoprecipitation of sRNAs with AGO1 or AGO2 from virus-infected plants revealed that p19 specifically impairs vsiRNA loading into AGO1 but not AGO2. Our findings, coupled with the fact that p19-expressing wild type Cymbidium ringspot virus (CymRSV) overcomes the Nicotiana benthamiana silencing based defense killing the host, suggest that AGO1 is the main effector of antiviral silencing in this host-virus combination. To further support our hypothesis we investigate whether the ability of p19 to bind endogenous sRNA without virus infection has biological important impact on endogenous pathways and is this reduced if the virus is present. To asses this we made mRNA sequencing from mock inoculated and Cym19stop infected p19syn plants. Cym19stop infected wild type plant was sequenced as a control. The sequencing data results supports our claims. An increase in transcriptional levels were found in case of genes known to be under small RNA regulation in uninfected p19syn plants and expressional levels return to normal Cym19stop p19syn plants.

Dataset Information

Proxitome profiling of a viral suppressors of RNA silencing identifies the DEAD-box RNA helicase RH20 as a positive regulator of antiviral RNA interference in Nicotiana benthamiana

Publications

DEAD-box RNA helicase RH20 positively regulates RNAi-based antiviral immunity in plants by associating with SGS3/RDR6 bodies.

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