The Arabidopsis F-box protein FBW2 targets AGO1 for degradation to prevent spurious loading of illegitimate small RNA

RNA silencing is a conserved mechanism in eukaryotes involved in development and defense against viruses. In plants, ARGONAUTE1 (AGO1) protein plays a central role in both microRNA- and small interfering RNA-directed silencing, and its expression is regulated at multiple levels. Here, we report that the F-box protein FBW2 assembles an SCF complex that selectively targets for proteolysis AGO1 when it is unloaded and mutated. Although FBW2 loss of function does not lead to strong growth or developmental defects, it significantly increases RNA-silencing activity. Interestingly, under conditions in which small-RNA accumulation is affected, the failure to degrade AGO1 in fbw2 mutants becomes more deleterious for the plant. Accordingly, the non-degradable AGO1 protein assembles high-molecular-weight complexes and binds illegitimate small RNA, leading to off-target cleavage. Therefore, control of AGO1 homeostasis by FBW2 plays an important role in quality control of RNA silencing. [Display omitted] •FBW2 assembles an SCF complex targeting specifically empty AGO1•FBW2 mutation leads to AGO1 accumulation and increased RNA silencing activity•Impaired sRNA accumulation in fbw2 results in AGO1 illegitimate loading•FBW2-mediated AGO1 proteolysis avoids off-target cleavage Argonaute proteins, main effectors of RNA silencing, remain stable only when they are loaded with small RNA. Hacquard et al. show that the E3 ubiquitin ligase component FBW2 triggers empty AGO1 degradation in Arabidopsis. Interfering with this mechanism leads to AGO1 spurious loading of illegitimate small RNA and off-target cleavage.