The RNA strands of the plus and minus polarities of peach latent mosaic viroid fold into different structures

Dubé, Audrey; Baumstark, Tilman; Bisaillon, Martin; Perreault, Jean‐Pierre

doi:10.1261/rna.1826710

Cited by 22 publications

(35 citation statements)

References 30 publications

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“…Different secondary structures between both polarities could explain why the exosome accesses more easily the (ϩ) polarity than the (Ϫ) polarity. Indeed, it has been shown that RNA strands of the (ϩ) and (Ϫ) polarities of PLMVd fold into two different structures (17). Furthermore, our results indicate that a fraction of the monomers are stabilized in the absence of the cytoplasmic 5Ј33Ј RNA degradation pathway.…”

Section: Discussionmentioning

confidence: 56%

Replication ofAvocado Sunblotch Viroidin the YeastSaccharomyces cerevisiae

Delan-Forino

Maurel

Torchet

2011

J Virol

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Viroids are the smallest known pathogenic agents. They are noncoding, single-stranded, closed-circular, "naked" RNAs, which replicate through RNA-RNA transcription. Viroids of the Avsunviroidae family possess a hammerhead ribozyme in their sequence, allowing self-cleavage during their replication. To date, viroids have only been detected in plant cells. Here, we investigate the replication of Avocado sunblotch viroid (ASBVd) of the Avsunviroidae family in a nonconventional host, the yeast Saccharomyces cerevisiae. We demonstrate that ASBVd RNA strands of both polarities are able to self-cleave and to replicate in a unicellular eukaryote cell. We show that the viroid monomeric RNA is destabilized by the nuclear 3 and the cytoplasmic 5 RNA degradation pathways. For the first time, our results provide evidence that viroids can replicate in other organisms than plants and that yeast contains all of the essential cellular elements for the replication of ASBVd.Viroids are the smallest and the simplest eukaryotic known pathogens (12). They are single-stranded circular RNAs capable of infecting plants and range from 246 to 401 nucleotides (nt). However, some variants of 120 or 475 nt are referenced in the subviral RNA database (47). Unlike viruses, they do not code for any protein, they are devoid of capsid or envelope, and are highly structured (13, 58). More than 34 viroid species have been described; they are divided into two families: the Pospiviroidae and the Avsunviroidae. Viroids of the Pospiviroidae family replicate via an RNA rolling-circle mechanism in the nucleus, and in the chloroplasts for the members of the Avsunviroidae family. Viroids move through the plant via the phloem and plasmodesmata that are part of the plant's vascular system and are transmitted by mechanical means, vegetative reproduction, and via seeds and insects (14,15,42). Members of the Avsunviroidae family such as Avocado sunblotch viroid (ASBVd) contain a catalytic hammerhead ribozyme and replicate through the following mechanism (20,21,53). The infectious monomeric circular RNA, which has positive-strand [(ϩ)] polarity, is transcribed by a host RNA polymerase, yielding oligomeric negative strands that self-cleave via the viroid ribozyme into monomers and circularize into negative-strand [(Ϫ)] circular viroids. These RNA species serve as templates to form (ϩ) polarity RNAs following the same steps in a symmetrical manner. According to this replication mechanism, the ribozyme undergoes self-cleavage on RNAs of both polarities. Therefore, both (ϩ) and (Ϫ) monomeric circular RNAs accumulate in tissues infected by Avsunviroidae.Based on these characteristics, ancestral viroids might have been present in an ancient RNA world (11, 23), one of the major scenarios of evolution in which the functional properties of nucleic acids and proteins observed today would have originally been performed by ancestral RNA molecules (2,30,37,41). In this context, it is interesting to explore the behavior of viroids and of other phylogenetically related satellit...

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Section: Discussionmentioning

confidence: 56%

Replication ofAvocado Sunblotch Viroidin the YeastSaccharomyces cerevisiae

Delan-Forino

Maurel

Torchet

2011

J Virol

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“…The experiment was also performed both with and without MgCl 2 and no pseudoknot was found in this structure (http://onlinelibrary.wiley.com/doi/10.1111/mpp.12130/suppinfo, see Supporting Information). This is in good agreement with previous experiments showing that the (–) strand is soluble in the presence of LiCl (Dube et al ., ), a property associated with rod‐like structures (Diener, ). The structure of the (–) strand variant of PLMVd.282 (NCBI accession number DQ680690) presented here corresponds relatively well with those reported previously for two variants (Dube et al ., ).…”

Section: Resultsmentioning

confidence: 99%

“…Prior to 2010, the secondary structures of only three viroid species had been solved in solution: PSTVd (Domdey et al ., ; Gast et al ., ; Gross et al ., ) and the Citrus exocortis viroid (CEVd) (Gross et al ., ) of the Pospiviroidae , and PLMVd (Bussiere et al ., ; Dube et al ., ) of the Avsunviroidae . Classic RNA probing approaches, such as RNAse treatment, are simply too laborious, and have not been adapted to probe RNA species the size of viroids.…”

Section: Introductionmentioning

confidence: 99%

Elucidation of the structures of all members of the Avsunviroidae family

Giguère

Adkar-Purushothama

Boyer

et al. 2014

Molecular Plant Pathology

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Viroids are small single-stranded RNA pathogens which cause significant damage to plants. As their nucleic acids do not encode for any proteins, they are dependant solely on their structure for their propagation. The elucidation of the secondary structures of viroids has been limited because of the exhaustive and time consuming nature of classic approaches. Here, the method of high-throughput selective 2'-hydroxyl acylation analysed by primer extension (hSHAPE) has been adapted to probe the viroid structure. The data obtained using this method were then used as input for computer-assisted structure prediction using RNA structure software in order to determine the secondary structures of the RNA strands of both (+) and (–) polarities of all Avsunviroidae members, one of the two families of viroids. The resolution of the structures of all of the members of the family provides a global view of the complexity of these RNAs. The structural differences between the two polarities, and any plausible tertiary interactions, were also analysed. Interestingly, the structures of the (+) and (–) strands were found to be different for each viroid species. The structures of the recently isolated grapevine hammerhead viroid-like RNA strands were also solved. This species shares several structural features with the Avsunviroidae family, although its infectious potential remains to be determined.To our knowledge, this article represents the first report of the structural elucidation of a complete family of viroids.

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“…The (+) and (−) polarities of PLMVd fold into different structures (Dubé et al, 2010). These have a branched secondary structure (Figure 5.2B) and do not have the conserved CCR, TCR, and TCH motifs but, remarkably, both their polarity strands self-cleave through hammerhead ribozymes (see Box 5.2).…”

Section: Nucleotide Sequencesmentioning

confidence: 99%

Agents Resembling or Altering Virus Diseases

Hull

2014

Plant Virology

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The RNA strands of the plus and minus polarities of peach latent mosaic viroid fold into different structures

Cited by 22 publications

References 30 publications

Replication ofAvocado Sunblotch Viroidin the YeastSaccharomyces cerevisiae

Replication ofAvocado Sunblotch Viroidin the YeastSaccharomyces cerevisiae

Elucidation of the structures of all members of the Avsunviroidae family

Agents Resembling or Altering Virus Diseases

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