Skip to Main content Skip to Navigation
Journal articles

Primer-Dependent and Primer-Independent Initiation of Double Stranded RNA Synthesis by Purified Arabidopsis RNA-Dependent RNA Polymerases RDR2 and RDR6

Abstract : Cellular RNA-dependent RNA polymerases (RDRs) are fundamental components of RNA silencing in plants and many other eukaryotes. In Arabidopsis thaliana genetic studies have demonstrated that RDR2 and RDR6 are involved in the synthesis of double stranded RNA (dsRNA) from single stranded RNA (ssRNA) targeted by RNA silencing. The dsRNA is subsequently cleaved by the ribonuclease DICER-like into secondary small interfering RNAs (siRNAs) that reinforce and/or maintain the silenced state of the target RNA. Models of RNA silencing propose that RDRs could use primer-independent and primer-dependent initiation to generate dsRNA from a transcript targeted by primary siRNA or microRNA (miRNA). However, the biochemical activities of RDR proteins are still partly understood. Here, we obtained active recombinant RDR2 and RDR6 in a purified form. We demonstrate that RDR2 and RDR6 have primer-independent and primer-dependent RNA polymerase activities with different efficiencies. We further show that RDR2 and RDR6 can initiate dsRNA synthesis either by elongation of 21-to 24-nucleotides RNAs hybridized to complementary RNA template or by elongation of self-primed RNA template. These findings provide new insights into our understanding of the molecular mechanisms of RNA silencing in plants.
Document type :
Journal articles
Complete list of metadatas

Cited literature [48 references]  Display  Hide  Download

https://hal-amu.archives-ouvertes.fr/hal-01217572
Contributor : Administrateur Hal Amu <>
Submitted on : Monday, October 19, 2015 - 5:44:10 PM
Last modification on : Tuesday, June 2, 2020 - 1:40:02 PM
Long-term archiving on: : Thursday, April 27, 2017 - 2:13:23 PM

File

fetchObject.pdf
Publisher files allowed on an open archive

Identifiers

Collections

Citation

Anthony Devert, Nicolas Fabre, Maïna Floris, Bruno Canard, Christophe Robaglia, et al.. Primer-Dependent and Primer-Independent Initiation of Double Stranded RNA Synthesis by Purified Arabidopsis RNA-Dependent RNA Polymerases RDR2 and RDR6. PLoS ONE, Public Library of Science, 2015, 10 (e0120100), ⟨10.1371/journal.pone.0120100.s006⟩. ⟨hal-01217572⟩

Share

Metrics

Record views

425

Files downloads

323