气候变化领域集成服务门户(CCPortal): Cooperative recruitment of RDR6 by SGS3 and SDE5 during small interfering RNA amplification in Arabidopsis

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DOI	10.1073/pnas.2102885118
	Cooperative recruitment of RDR6 by SGS3 and SDE5 during small interfering RNA amplification in Arabidopsis
	Yoshikawa M.; Han Y.-W.; Fujii H.; Aizawa S.; Nishino T.; Ishikawa M.
发表日期	2021
ISSN	0027-8424
卷号	118 期号:34
英文摘要	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 Arabidopsis thaliana 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. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	Argonaute; RNA silencing; RNA-dependent RNA polymerase; Secondary siRNA; SiRNA
语种	英语
scopus关键词	Arabidopsis protein; double stranded RNA; RDR6 protein, Arabidopsis; RNA induced silencing complex; SDE5 protein, Arabidopsis; SGS3 protein, Arabidopsis; small interfering RNA; Arabidopsis; gene expression regulation; genetics; metabolism; Arabidopsis; Arabidopsis Proteins; Gene Expression Regulation, Plant; RNA, Double-Stranded; RNA, Small Interfering; RNA-Dependent RNA Polymerase; RNA-Induced Silencing Complex
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/251041
作者单位	Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, Ibaraki, Tsukuba, 305-8518, Japan; Laboratory for Integrative Genomics, RIKEN Center for Integrative Medical Sciences, Yokohama, 230-0045, Japan; Faculty of Advanced Engineering, Tokyo University of Science, Katsushika-ku, Tokyo, 125-8585, Japan
推荐引用方式 GB/T 7714	Yoshikawa M.,Han Y.-W.,Fujii H.,et al. Cooperative recruitment of RDR6 by SGS3 and SDE5 during small interfering RNA amplification in Arabidopsis[J],2021,118(34).
APA	Yoshikawa M.,Han Y.-W.,Fujii H.,Aizawa S.,Nishino T.,&Ishikawa M..(2021).Cooperative recruitment of RDR6 by SGS3 and SDE5 during small interfering RNA amplification in Arabidopsis.Proceedings of the National Academy of Sciences of the United States of America,118(34).
MLA	Yoshikawa M.,et al."Cooperative recruitment of RDR6 by SGS3 and SDE5 during small interfering RNA amplification in Arabidopsis".Proceedings of the National Academy of Sciences of the United States of America 118.34(2021).