The RYMV-Encoded Viral Suppressor of RNA Silencing P1 Is a Zinc-Binding Protein with Redox-Dependent Flexibility

Gillet, François-Xavier; Cattoni, Diego I.; Petiot-Bécard, Stéphanie; Delalande, François; Poignavent, Vianney; Brizard, Jean-Paul; Bessin, Yannick; Dorsselaer, Alain Van; Declerck, Nathalie; Sanglier-Cianférani, Sarah; Brugidou, Christophe; Vignols, Florence

doi:10.1016/j.jmb.2013.03.028

Cited by 23 publications

(40 citation statements)

References 68 publications

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“…Indeed, these two characteristics are possibly involved in silencing suppression mechanisms through a nucleic acid binding function [41][42][43]. In addition, candidate proteins were also selected for the presence of WG/GW motifs (the nucleotide sequence coding for Glycine-G and Tryptophan-W), which have been demonstrated to be involved in the suppression of silencing via AGO1 sequestration [44].…”

Section: Selection Of Vsr Candidate Proteinsmentioning

confidence: 99%

p2 of Rice grassy stunt virus (RGSV) and p6 and p9 of Rice ragged stunt virus (RRSV) isolates from Vietnam exert suppressor activity on the RNA silencing pathway

et al. 2015

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In Vietnam, the two main viruses that cause disease in rice are the Rice grassy stunt virus (RGSV) and the Rice ragged stunt virus (RRSV). Outbreaks of these two viruses have dramatically decreased rice production in Vietnam. Because natural resistance genes are unknown, an RNAi strategy may be an alternative method to develop resistance to RGSV and RRSV. However, this strategy will be efficient only if putative silencing suppressors encoded by the two viruses are neutralized. To identify these suppressors, we used the classical green fluorescent protein (GFP) agroinfiltration method in Nicotiana benthamiana. Then, we investigated the effects of viral candidate proteins on GFP expression and GFP siRNA accumulation and their interference with the short- or long-range signal of silencing. RGSV genes s2gp1, s5gp2, and s6gp1 and RRSV genes s5gp1, s6gp1, s9gp1, and s10gp1 were selected for viral silencing suppressor investigation according to their small molecular weight, the presence of cysteines, or the presence of a GW motif in related protein products. We confirmed that protein p6 of RRSV displays mild silencing suppressor activity and affects long-range silencing by delaying the systemic silencing signal. In addition, we identified two new silencing suppressors that displayed mild activity: p2 of RGSV and p9 of RRSV.

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Section: Selection Of Vsr Candidate Proteinsmentioning

confidence: 99%

p2 of Rice grassy stunt virus (RGSV) and p6 and p9 of Rice ragged stunt virus (RRSV) isolates from Vietnam exert suppressor activity on the RNA silencing pathway

et al. 2015

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“…Our previous analyses suggested that P1 binds its two zinc atoms with differential affinity (Gillet et al, 2013). To further address this question, we first performed EDTA treatments followed by native ESI-MS analyses on the full-length P1 protein and found that treatments up to 400 molar equivalents (eq) of EDTA were sufficient to provoke the release of one zinc atom (Table 1 and Appendix Fig S6A, compare upper and mid panels).…”

Section: Znf1 and Znf2 In P1 Exhibit Different Zinc Binding Affinitymentioning

confidence: 97%

“…In a previous work, we found that P1 binds two zinc atoms in a reduced environment (Gillet et al, 2013), but the high number of rigorously conserved cysteine (7) and histidine (6) residues in P1 diversity (Appendix Fig S1) prevented the identification of the amino acids involved. To address this question, we designed a series of recombinant P1 regions ( Fig Table 1-Table 2) and evaluated their capacity to bind zinc using a 4-(2-Pyridylazo) resorcinol (PAR) labeling assays.…”

Section: Delineation Of P1 Zinc Binding Domainsmentioning

confidence: 98%

“…Point mutations were introduced on Cys residue of each Cys-X2-Cys motif (Cys 64 and Cys 95), which impaired silencing suppression and cell-to-cell movement in transgenic rice respectively, indicating a central role of Cys residues for P1 functions (Siré et al, 2008). Using a biochemical approach coupled with mass spectrometry, we showed that P1 binds two Zn atoms in its reduced state, whereas its oxidation induced complete Zn release accompanied by disulfide bonds formation, structural modification and oligomerization (Gillet et al, 2013). This process could be reverted by adding a reducing agent, indicating that P1 is a Zn binding protein with redox flexibility and oligomerization properties in vitro, highlighting the importance of Cys residues for P1 behavior.…”

Section: Introductionmentioning

confidence: 99%

“…All functions achieved by P1 play a crucial role during early steps of viral infection, however little is known concerning its mode of action. P1 presents two couples of Cysteine (Cys) organized in Cys-X2-Cys motifs similar to a zinc finger (ZnF) (Gillet et al, 2013) and highly conserved among RYMV P1 diversity (Cys 64-X2-Cys 67 and Cys 92-X2-Cys 95), and remarkably, in the P1 of sobemovirus genus (Sõmera et al, 2015). Point mutations were introduced on Cys residue of each Cys-X2-Cys motif (Cys 64 and Cys 95), which impaired silencing suppression and cell-to-cell movement in transgenic rice respectively, indicating a central role of Cys residues for P1 functions (Siré et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

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An original structural fold underlies the multitask P1, a silencing suppressor encoded by the Rice yellow mottle virus

Poignavent

Hoh

Terral

et al. 2020

Preprint

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The Rice Yellow Mottle sobemovirus (RYMV) belongs to the most damaging pathogens devastating rice fields in Africa. P1, a key protein for RYMV, was reported as a potent RNAi suppressor counteracting RNA silencing in plant reporter systems. Here we describe the complete 3D structure and dynamics of P1. Its N-terminal region contains ZnF1, a structural CCCC-type zinc finger strongly affine to zinc and a prominent short helix, rendering this region poorly amenable to structural changes. P1 C-terminal region contains ZnF2, an atypical HCHC-type ZnF that does not belong to any existing class of Zn finger proteins. ZnF2 appeared much less affine to zinc and more sensitive to oxidizing environments than ZnF1, and may serve as a sensor of plant redox status. We also identified key residues essential for RYMV infectivity and spread in rice tissues through their participation in P1 oligomerization and folding.Altogether, our results provide the first complete structure of a rice antiviral silencing suppressor and highlight P1 structural properties that may serve RYMV functions to infect and invade the host plant.

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Complete nucleotide sequence of Solanum nodiflorum mottle virus

Sõmera

Truve

2017

Arch Virol

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Solanum nodiflorum mottle virus (SNMoV) was isolated from a small-flowered nightshade (Solanum nodiflorum) in Queensland, Australia. It has been included in the genus Sobemovirus based on virion morphology and serological relationships. Here, we report the sequence of the complete genome of SNMoV. Sequence analysis confirmed that SNMoV has the characteristic genome organization of sobemoviruses. Phylogenetic analysis showed that it clusters most closely with velvet tobacco mottle virus (VTMoV), another sobemovirus native to Australia. Their genomes show 56.8 % sequence identity.

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The RYMV-Encoded Viral Suppressor of RNA Silencing P1 Is a Zinc-Binding Protein with Redox-Dependent Flexibility

Cited by 23 publications

References 68 publications

p2 of Rice grassy stunt virus (RGSV) and p6 and p9 of Rice ragged stunt virus (RRSV) isolates from Vietnam exert suppressor activity on the RNA silencing pathway

p2 of Rice grassy stunt virus (RGSV) and p6 and p9 of Rice ragged stunt virus (RRSV) isolates from Vietnam exert suppressor activity on the RNA silencing pathway

An original structural fold underlies the multitask P1, a silencing suppressor encoded by the Rice yellow mottle virus

Complete nucleotide sequence of Solanum nodiflorum mottle virus

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