RDR2 Partially Antagonizes the Production of RDR6-Dependent siRNA in Sense Transgene-Mediated PTGS

Jauvion, Vincent; Rivard, Maud; Bouteiller, Nathalie; Elmayan, Taline; Vaucheret, Hervé

doi:10.1371/journal.pone.0029785

Cited by 34 publications

(23 citation statements)

References 28 publications

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“…In the slicer-deficient ago1 background, TAS1c siRNAs remained unaffected by inactivation of RDR1 ( Figure 5D). RDR2 is mainly involved in the biosynthesis of 24-nucleotide siRNAs that guide transcriptional gene silencing (Xie et al, 2004), but it can also influence the production of other small RNAs (Jauvion et al, 2012;Klein-Cosson et al, 2015). Mutation of RDR2 had only a small effect on TAS1c siRNA accumulation in the slicer-deficient ago1 background ( Figure 5E).…”

Section: Dependence On Rdrs and Sgs3 Of Unphased Tasirnasmentioning

confidence: 99%

The Slicer Activity of ARGONAUTE1 is Required Specifically for the Phasing, Not Production, of Trans-Acting Short Interfering RNAs in Arabidopsis

et al. 2016

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ARGONAUTE1 (AGO1) mediates posttranscriptional silencing by microRNAs (miRNAs) and short interfering RNAS (siRNAs). AGO1-catalyzed RNA cleavage (slicing) represses miRNA targets, but current models also highlight the roles of slicing in formation of siRNAs and siRNA-AGO1 complexes. miRNA-guided slicing is required for biogenesis of phased, trans-acting siRNAs (tasiRNAs), whose cleaved precursor fragments are converted to double-stranded RNA by RNA-dependent RNA polymerase 6 (RDR6). In addition, unwinding of duplex siRNA bound to AGO1 requires passenger strand cleavage in vitro. In this study, we analyze how mutation of four metal ion-coordinating residues of Arabidopsis thaliana AGO1 affects slicer activity in vitro and siRNA function in vivo. We show that while all four residues are required for slicer activity, they do not contribute equally to catalysis. Moreover, passenger strand cleavage is required for assembly of active AGO1-siRNA complexes in vivo, and many AGO1-bound siRNAs are trimmed in the absence of slicer activity. Remarkably, seedlings defective in AGO1 slicer activity produce abundant siRNAs from tasiRNA loci in vivo. These siRNAs depend on RDR6 and SUPPRESSOR OF GENE SILENCING3, but unlike wild-type tasiRNAs, they are unphased. These results demonstrate that slicing is solely required for phase definition of tasiRNAs, and they strongly support recruitment of RDR6 by AGO1 rather than by cleavage fragments.

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Section: Dependence On Rdrs and Sgs3 Of Unphased Tasirnasmentioning

confidence: 99%

The Slicer Activity of ARGONAUTE1 is Required Specifically for the Phasing, Not Production, of Trans-Acting Short Interfering RNAs in Arabidopsis

et al. 2016

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“…24nt and 21nt siRNA pathways antagonize each other in plants (Creasey et al, 2014; Jauvion et al, 2012; Mari-Ordonez et al, 2013), reminiscent of WAGO and CSR-1 22Gs in C.elegans , and may participate in scanning for “non-self” transposons in pollen (Slotkin et al, 2009). 21nt secondary siRNA (epigenetically activated siRNA or easiRNA) are triggered by miRNA (Creasey et al, 2014), and target transposons that are strongly activated in the vegetative nucleus (Figure 2), accompanied by down regulation of DDM1 (Calarco et al, 2012; Ibarra et al, 2012; Slotkin et al, 2009).…”

Section: Germline Reprogramming and Imprintingmentioning

confidence: 99%

Transgenerational Epigenetic Inheritance: Myths and Mechanisms

Heard

Martienssen

2014

Cell

1,450

1,151

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Since the human genome was sequenced, the term “epigenetics” is increasingly being associated with the hope that we are more than just the sum of our genes. Might what we eat, the air we breathe, or even the emotions we feel, influence not only our genes but those of descendents? The environment can certainly influence gene expression, and can lead to disease, but trans-generational consequences are another matter. While the inheritance of epigenetic characters can certainly occur - particularly in plants – how much is due to the environment and the extent to which it happens in humans, remains unclear.

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“…RDR1 and RDR6 have redundant functions in the biosynthesis or amplification of viral dsRNAs for TuMV and CMV (Garcia-Ruiz et al, 2010, Wang et al, 2010. RDR2 and RDR6 compete for RNA substrates produced by transgenes that are subjected to sense transgenemediated posttranscriptional gene silencing (S-PTGS) (Jauvion et al, 2012). Data from the previous and present studies indicates that RDR1, RDR2, and RDR6 have redundant functions in the biosynthesis of antisense RNAs from sense transcripts of protein-coding genes.…”

Section: Discussionmentioning

confidence: 69%

Novel Stress-Inducible Antisense RNAs of Protein-Coding Loci Are Synthesized by RNA-Dependent RNA Polymerase

et al. 2017

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(S.S.); 0000-0001-8288-0467 (M.S.).Our previous study identified approximately 6,000 abiotic stress-responsive noncoding transcripts existing on the antisense strand of protein-coding genes and implied that a type of antisense RNA was synthesized from a sense RNA template by RNAdependent RNA polymerase (RDR). Expression analyses revealed that the expression of novel abiotic stress-induced antisense RNA on 1,136 gene loci was reduced in the rdr1/2/6 mutants. RNase protection indicated that the RD29A antisense RNA and other RDR1/2/6-dependent antisense RNAs are involved in the formation of dsRNA. The accumulation of stress-inducible antisense RNA was decreased and increased in dcp5 and xrn4, respectively, but not changed in dcl2/3/4, nrpd1a and nrpd1b. RNA-seq analyses revealed that the majority of the RDR1/2/6-dependent antisense RNA loci did not overlap with RDR1/2/6-dependent 20-30 nt RNA loci. Additionally, rdr1/2/6 mutants decreased the degradation rate of the sense RNA and exhibited arrested root growth during the recovery stage following a drought stress, whereas dcl2/3/4 mutants did not. Collectively, these results indicate that RDRs have stress-inducible antisense RNA synthesis activity and a novel biological function that is different from the known endogenous small RNA pathways from protein-coding genes. These data reveal a novel mechanism of RNA regulation during abiotic stress response that involves complex RNA degradation pathways.

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RDR2 Partially Antagonizes the Production of RDR6-Dependent siRNA in Sense Transgene-Mediated PTGS

Cited by 34 publications

References 28 publications

The Slicer Activity of ARGONAUTE1 is Required Specifically for the Phasing, Not Production, of Trans-Acting Short Interfering RNAs in Arabidopsis

The Slicer Activity of ARGONAUTE1 is Required Specifically for the Phasing, Not Production, of Trans-Acting Short Interfering RNAs in Arabidopsis

Transgenerational Epigenetic Inheritance: Myths and Mechanisms

Novel Stress-Inducible Antisense RNAs of Protein-Coding Loci Are Synthesized by RNA-Dependent RNA Polymerase

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