<i>Retracted:</i> Identification, subcellular localization and some properties of a cysteine‐rich suppressor of gene silencing encoded by peanut clump virus

Dunoyer, Patrice; Pfeffer, Sébastien; Fritsch, C.; Hemmer, O.; Voinnet, Olivier; Richards, K.

doi:10.1046/j.0960-7412.2001.01242.x

Cited by 132 publications

(70 citation statements)

References 79 publications

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“…Inserts were then digested and cloned into the XbaI and BamHI sites of pBIN61-FLAG. 35S: HA-AGO2 and 35S:HA-AGO5 slicer-defective variants were generated by PCR mutagenesis using pBIC-HA-AGO2 and pBIC-HA-AGO5 as templates with primers listed in Supplemental Table 1. PVX, PVX-GFP, and PVX-GFPDTGB binary constructs Bhattacharjee et al, 2009), PlAMV-GFP (Yamaji et al, 2012), 35S: P14, 35S:P21 (Mérai et al, 2006), 35S:P15 (Dunoyer et al, 2002), and 35:P19 have been previously described. pBIN61-P25:HA was generated by RT-PCR from PVX-infected plants using primers (Supplemental Table 1) to introduce XbaI and BamHI sites at the 59 and 39 ends of the P25 open reading frame, respectively.…”

Section: Plasmid Constructionmentioning

confidence: 99%

Functional and Genetic Analysis Identify a Role for Arabidopsis ARGONAUTE5 in Antiviral RNA Silencing

2015

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RNA silencing functions as an antiviral defense through the action of DICER-like (DCL) and ARGONAUTE (AGO) proteins. In turn, plant viruses have evolved strategies to counteract this defense mechanism, including the expression of suppressors of RNA silencing. Potato virus X (PVX) does not systemically infect Arabidopsis thaliana Columbia-0, but is able to do so effectively in mutants lacking at least two of the four Arabidopsis DCL proteins. PVX can also infect Arabidopsis ago2 mutants, albeit less effectively than double DCL mutants, suggesting that additional AGO proteins may mediate anti-viral defenses. Here we show, using functional assays, that all Arabidopsis AGO proteins have the potential to target PVX lacking its viral suppressor of RNA silencing (VSR), P25, but that only AGO2 and AGO5 are able to target wild-type PVX. However, P25 directly affects only a small subset of AGO proteins, and we present evidence indicating that its protective effect is mediated by precluding AGO proteins from accessing viral RNA, as well as by directly inhibiting the RNA silencing machinery. In agreement with functional assays, we show that Potexvirus infection induces AGO5 expression and that both AGO2 and AGO5 are required for full restriction of PVX infection in systemic tissues of Arabidopsis.

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Section: Plasmid Constructionmentioning

confidence: 99%

Functional and Genetic Analysis Identify a Role for Arabidopsis ARGONAUTE5 in Antiviral RNA Silencing

2015

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“…Among the best-studied examples are the helper component-proteinase (HC-Pro) of the potyvirus Potato virus Y (PVY) and the 2b protein of Cucumber mosaic virus (CMV) (7,40). Other plus-strand RNA (and some DNA) viruses also have been found to suppress gene silencing, and for some of them the viral protein involved was identified (16,37,46,54). The viral suppressor proteins of PVY and CMV act differently by targeting different steps in the RNA silencing pathway.…”

mentioning

confidence: 99%

Negative-Strand Tospoviruses and Tenuiviruses Carry a Gene for a Suppressor of Gene Silencing at Analogous Genomic Positions

Bucher¹,

Sijen

Haan³

et al. 2003

J Virol

219

166

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Posttranscriptional silencing of a green fluorescent protein (GFP) transgene in Nicotiana benthamiana plants was suppressed when these plants were infected with Tomato spotted wilt virus (TSWV), a plant-infecting member of the Bunyaviridae. Infection with TSWV resulted in complete reactivation of GFP expression, similar to the case for Potato virus Y, but distinct from that for Cucumber mosaic virus, two viruses known to carry genes encoding silencing suppressor proteins. Agrobacterium-based leaf injections with individual TSWV genes identified the NS S gene to be responsible for the RNA silencing-suppressing activity displayed by this virus. The absence of short interfering RNAs in NS S -expressing leaf sectors suggests that the tospoviral NS S protein interferes with the intrinsic RNA silencing present in plants. Suppression of RNA silencing was also observed when the NS3 protein of the Rice hoja blanca tenuivirus, a nonenveloped negative-strand virus, was expressed. These results indicate that plant-infecting negative-strand RNA viruses carry a gene for a suppressor of RNA silencing.RNA silencing involves a sequence-specific degradation which is induced by overabundant RNA and by doublestranded RNA (dsRNA) molecules and which can target transgenes as well as homologous endogenous genes. RNA silencing was first described for plants (35,50) and over recent years has been described for other organisms, where it is also referred to as cosuppression, posttranscriptional gene silencing (17), or RNA-mediated virus resistance (3,11,30) in plants, quelling in fungi (9), or RNAi in animals (19). Building blocks of the gene-silencing pathway proved to have remarkable similarities in the different organisms and hence suggest an ancient role of gene silencing in pathogen resistance or development (10,25,53). One of the key intermediary elements in the RNA silencing pathway is dsRNA, which is recognized by a dsRNAspecific nuclease (5) to yield small (21 to 23 nucleotides) short interfering RNAs (siRNAs) (21). These siRNAs subsequently serve as guides for cleavage of homologous RNA molecules. In plants, versions of transgenes that produce dsRNA molecules have been shown to be very potent activators of RNA silencing (47). As all RNA viruses replicate through formation of dsRNA intermediates, these are potential targets of the RNA silencing mechanism. Indeed, antiviral RNA silencing has been shown to occur in nature and has been proposed as a natural defense mechanism protecting plants against viruses, resulting in resistance (1, 43).To counteract the RNA silencing mechanism of their host, plant viruses have developed ways to evade or neutralize this response. Over recent years, RNA silencing-inhibiting proteins have been identified in several plant viruses. Among the best-studied examples are the helper component-proteinase (HC-Pro) of the potyvirus Potato virus Y (PVY) and the 2b protein of Cucumber mosaic virus (CMV) (7, 40). Other plus-strand RNA (and some DNA) viruses also have been found to suppress gene silencing, and ...

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“…Infiltration of non-transgenic Nicotiana benthamiana leaves with a strain of Agrobacterium tumefaciens carrying a gfp gene construct (35S-gfp) results in transient GFP expression, observed as a green fluorescence under UV illumination (Figure 1a) (Llave et al 2000;Voinnet et al 2000;Dunoyer et al 2002;Hamilton et al 2002;Takeda et al 2002;Bucher et al 2003). In contrast, co-infiltration of agrobacteria carrying a 35S-gfp construct together with a strain carrying an inverted repeat of gfp construct (35S-ds gfp) does not produce any green fluorescence.…”

Section: Gene Silencing Of Transiently Expressed Sequencesmentioning

confidence: 99%

A practical approach to the understanding and teaching of RNA silencing in plants

Bazzini

Mongelli

Hopp

et al. 2007

Electron. J. Biotechnol.

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Retracted: Identification, subcellular localization and some properties of a cysteine‐rich suppressor of gene silencing encoded by peanut clump virus

Cited by 132 publications

References 79 publications

Functional and Genetic Analysis Identify a Role for Arabidopsis ARGONAUTE5 in Antiviral RNA Silencing

Functional and Genetic Analysis Identify a Role for Arabidopsis ARGONAUTE5 in Antiviral RNA Silencing

Negative-Strand Tospoviruses and Tenuiviruses Carry a Gene for a Suppressor of Gene Silencing at Analogous Genomic Positions

A practical approach to the understanding and teaching of RNA silencing in plants

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