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Cooperative recruitment of RDR6 by SGS3 and SDE5 during small interfering RNA amplification in Arabidopsis

Proceedings of the National Academy of Sciences - PNAS, 2021-08, Vol.118 (34), p.1 [Peer Reviewed Journal]

Copyright National Academy of Sciences Aug 24, 2021 ;2021 ;ISSN: 0027-8424 ;EISSN: 1091-6490 ;DOI: 10.1073/pnas.2102885118 ;PMID: 34408020

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Title:
Cooperative recruitment of RDR6 by SGS3 and SDE5 during small interfering RNA amplification in Arabidopsis
Author: Yoshikawa, Manabu ; Han, Yong-Woon ; Fujii, Hirofumi ; Aizawa, Shu ; Nishino, Tatsuya ; Ishikawa, Masayuki
Subjects: Amplification ; Arabidopsis - genetics ; Arabidopsis - metabolism ; Arabidopsis Proteins - genetics ; Arabidopsis Proteins - metabolism ; Biological Sciences ; Cleavage ; DNA-directed RNA polymerase ; Double-stranded RNA ; Gene Expression Regulation, Plant ; Gene silencing ; Nucleotides ; Poly(A) ; Polyadenylation ; Proteins ; Recruitment ; Ribonucleic acid ; RNA ; RNA polymerase ; RNA, Double-Stranded - genetics ; RNA, Double-Stranded - metabolism ; RNA, Small Interfering - genetics ; RNA, Small Interfering - metabolism ; RNA-Dependent RNA Polymerase - genetics ; RNA-Dependent RNA Polymerase - metabolism ; RNA-directed RNA polymerase ; RNA-Induced Silencing Complex - genetics ; RNA-Induced Silencing Complex - metabolism ; RNA-mediated interference ; siRNA
Is Part Of: Proceedings of the National Academy of Sciences - PNAS, 2021-08, Vol.118 (34), p.1
Description: Small interfering RNAs (siRNAs) are often amplified from transcripts cleaved by RNA-induced silencing complexes (RISCs) containing a small RNA (sRNA) and an Argonaute protein. Amplified siRNAs, termed secondary siRNAs, are important for reinforcement of target repression. In plants, target cleavage by RISCs containing 22-nucleotide (nt) sRNA and Argonaute 1 (AGO1) triggers siRNA amplification. In this pathway, the cleavage fragment is converted into double-stranded RNA (dsRNA) by RNA-dependent RNA polymerase 6 (RDR6), and the dsRNA is processed into siRNAs by Dicer-like proteins. Because nonspecific RDR6 recruitment causes nontarget siRNA production, it is critical that RDR6 is specifically recruited to the target RNA that serves as a template for dsRNA formation. Previous studies showed that Suppressor of Gene Silencing 3 (SGS3) binds and stabilizes 22-nt sRNA-containing AGO1 RISCs associated with cleaved target, but how RDR6 is recruited to targets cleaved by 22-nt sRNA-containing AGO1 RISCs remains unknown. Here, using cell-free extracts prepared from suspension-cultured cells, we established an in vitro system for secondary siRNA production in which 22-nt siRNA-containing AGO1-RISCs but not 21-nt siRNA-containing AGO1-RISCs induce secondary siRNA production. In this system, addition of recombinant Silencing Defective 5 (SDE5) protein remarkably enhances secondary siRNA production. We show that RDR6 is recruited to a cleavage fragment by 22-nt siRNA-containing AGO1-RISCs in coordination with SGS3 and SDE5. The SGS3-SDE5-RDR6 multicomponent recognition system and the poly(A) tail inhibition may contribute to securing specificity of siRNA amplification.
Publisher: United States: National Academy of Sciences
Language: English
Identifier: ISSN: 0027-8424
EISSN: 1091-6490
DOI: 10.1073/pnas.2102885118
PMID: 34408020
Source: GFMER Free Medical Journals
MEDLINE
PubMed Central

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Cooperative recruitment of RDR6 by SGS3 and SDE5 during small interfering RNA amplification in Arabidopsis

Copyright National Academy of Sciences Aug 24, 2021 ;2021 ;ISSN: 0027-8424 ;EISSN: 1091-6490 ;DOI: 10.1073/pnas.2102885118 ;PMID: 34408020

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