Phase separation of SGS3 drives siRNA body formation and promotes endogenous gene silencing

Tan, Huijuan1,2,3; Luo, Wentao1,2; Yan, Wei1,2; Liu, Jie4; Aizezi, Yalikunjiang1,2; Cui, Ruixue5; Tian, Ruijun4; Ma, Jinbiao5; Guo, Hongwei1,2

2023-01-31

10.1016/j.celrep.2022.111985

Cell Reports   Impact Factor & Quartile

2211-1247

The generation of small interfering RNA (siRNA) involves many RNA processing components, including SUP-PRESSOR OF GENE SILENCING 3 (SGS3), RNA-DEPENDENT RNA POLYMERASE 6 (RDR6), and DICER -LIKE proteins (DCLs). Nonetheless, how these components are coordinated to produce siRNAs is unclear. Here, we show that SGS3 forms condensates via phase separation in vivo and in vitro. SGS3 interacts with RDR6 and drives it to form siRNA bodies in cytoplasm, which is promoted by SGS3-targeted RNAs. Disrupt-ing SGS3 phase separation abrogates siRNA body assembly and siRNA biogenesis, whereas coexpression of SGS3 and RDR6 induces siRNA body formation in tobacco and yeast cells. Dysfunction in translation and mRNA decay increases the number of siRNA bodies, whereas DCL2/4 mutations enhance their size. Purifi-cation of SGS3 condensates identifies numerous RNA-binding proteins and siRNA processing components. Together, our findings reveal that SGS3 phase separation-mediated formation of siRNA bodies is essential for siRNA production and gene silencing.

SCI

English

First ; Corresponding

National Key R&D Program of China[91940303] ; National Natural Science Foundation of China["31900459","2018YFA0507101"] ; National Key Research and Development Program of China[20200925161843002] ; null[2018YFA0507100]

Cell Biology

Cell Biology

WOS:000925296600001

CELL PRESS

Web of Science

1.Southern Univ Sci & Technol, Sch Life Sci, lInst Plant & Food Sci, Dept Biol, Shenzhen 518055, Guangdong, Peoples R China
2.Southern Univ Sci & Technol, Lab Mol Design Plant Cell Factory, Guangdong Higher Educ Inst, Shenzhen 518055, Peoples R China
3.Wuhan Univ, Inst Adv Studies, Wuhan 430072, Peoples R China
4.Southern Univ Sci & Technol, Coll Sci, Dept Chem, Shenzhen 518055, Peoples R China
5.Fudan Univ, Huashan Hosp,State Key Lab Genet Engn, Multiscale Res Inst Complex Syst,Dept Biochem & B, Collaborat Innovat Ctr Genet & Dev,Sch Life Sci, Shanghai 200438, Peoples R China

Tan, Huijuan,Luo, Wentao,Yan, Wei,et al. Phase separation of SGS3 drives siRNA body formation and promotes endogenous gene silencing[J]. Cell Reports,2023,42(1).

Tan, Huijuan.,Luo, Wentao.,Yan, Wei.,Liu, Jie.,Aizezi, Yalikunjiang.,...&Guo, Hongwei.(2023).Phase separation of SGS3 drives siRNA body formation and promotes endogenous gene silencing.Cell Reports,42(1).

Tan, Huijuan,et al."Phase separation of SGS3 drives siRNA body formation and promotes endogenous gene silencing".Cell Reports 42.1(2023).