Journal
EMBO JOURNAL
Volume 39, Issue 20, Pages -Publisher
WILEY
DOI: 10.15252/embj.2020105130
Keywords
Ago3; nuage; phosphorylation; Siwi; Vreteno
Categories
Funding
- Ministry of Education, Culture, Sports, Science and Technology (MEXT) [19H05466, 20K06483]
- Japan Society for the Promotion of Science
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Silkworm ovarian germ cells produce the Siwi-piRNA-induced silencing complex (piRISC) through two consecutive mechanisms, the primary pathway and the secondary ping-pong cycle. Primary Siwi-piRISCproduction occurs on the outer mitochondrial membrane in an Ago3-independent manner, where Tudor domain-containing Papi binds unloaded Siwi via its symmetrical dimethylarginines (sDMAs). Here, we now show that secondary Siwi-piRISCproduction occurs at the Ago3-positive nuage Ago3 bodies, in an Ago3-dependent manner, where Vreteno (Vret), another Tudor protein, interconnects unloaded Siwi and Ago3-piRISCthrough theirsDMAs. Upon Siwi depletion, Ago3 is phosphorylated and insolubilized in its piRISCform with cleavedRNAs and Vret, suggesting that the complex is stalled in the intermediate state. The Ago3 bodies are also enlarged. The aberrant morphology is restored upon Siwi re-expression without Ago3-piRISCsupply. Thus, Siwi depletion aggregates the Ago3 bodies to protect the piRNAintermediates from degradation until the normal cellular environment returns to re-initiate the ping-pong cycle. Overall, these findings reveal a unique regulatory mechanism controlling piRNAbiogenesis.
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