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The Arabidopsis F-box protein FBW2 degrades AGO1 to avoid spurious loading of illegitimate small RNA [PARE-seq]

ABSTRACT: RNA silencing is a conserved mechanism in eukaryotes and is involved in development, heterochromatin maintenance and defense against viruses. In plants, ARGONAUTE1 (AGO1) protein plays a central role in both microRNA (miRNA) and small interfering RNA (siRNA)-directed silencing and its expression is regulated at multiple levels. Here we report that the F-box protein FBW2 targets proteolysis of AGO1 by a CDC48-mediated autophagy mechanism. We found that FBW2 assembles an SCF complex that recognizes the MID-PIWI domain of AGO1 and requires its C-terminal domain containing a GW motif for AGO1 turnover. We show that FBW2 has a preference for unloaded AGO1 and also for some mutated forms of the protein. FBW2 loss of function increases RNA silencing activity, but does not lead to strong growth or developmental defects. However, under conditions in which small RNA production or accumulation is affected, the failure to degrade AGO1 in fbw2 mutants becomes more deleterious for the plant. Hence, we show that the non-degradable AGO1 protein assembles high molecular complexes and binds illegitimate small RNAs.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE169433 | GEO | 2022/04/01

REPOSITORIES: GEO

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The Arabidopsis F-box protein FBW2 degrades AGO1 to avoid spurious loading of illegitimate small RNA [ncRNA-Seq]

Project description:RNA silencing is a conserved mechanism in eukaryotes and is involved in development, heterochromatin maintenance and defense against viruses. In plants, ARGONAUTE1 (AGO1) protein plays a central role in both microRNA (miRNA) and small interfering RNA (siRNA)-directed silencing and its expression is regulated at multiple levels. Here we report that the F-box protein FBW2 targets proteolysis of AGO1 by a CDC48-mediated autophagy mechanism. We found that FBW2 assembles an SCF complex that recognizes the MID-PIWI domain of AGO1 and requires its C-terminal domain containing a GW motif for AGO1 turnover. We show that FBW2 has a preference for unloaded AGO1 and also for some mutated forms of the protein. FBW2 loss of function increases RNA silencing activity, but does not lead to strong growth or developmental defects. However, under conditions in which small RNA production or accumulation is affected, the failure to degrade AGO1 in fbw2 mutants becomes more deleterious for the plant. Hence, we show that the non-degradable AGO1 protein assembles high molecular complexes and binds illegitimate small RNAs.

2022-04-01 | GSE169324 | GEO

The Arabidopsis F-box protein FBW2 degrades AGO1 to avoid spurious loading of illegitimate small RNA

Project description:RNA silencing is a conserved mechanism in eukaryotes and is involved in development, heterochromatin maintenance and defense against viruses. In plants, ARGONAUTE1 (AGO1) protein plays a central role in both microRNA (miRNA) and small interfering RNA (siRNA)-directed silencing and its expression is regulated at multiple levels. Here, we report that the F-box protein FBW2 targets proteolysis of AGO1 by a CDC48-mediated mechanism. We found that FBW2 assembles an SCF complex that recognizes the MID-PIWI domain of AGO1 and requires its C-terminal domain containing a GW motif for AGO1 turnover. We showed that FBW2 has a preference for the unloaded and for some mutated forms of AGO1 protein. While 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 production or accumulation is affected, the failure to degrade AGO1 in fbw2 mutants becomes more deleterious for the plant. Hence, we showed that the non-degradable AGO1 protein assembles high molecular complexes and binds illegitimate small RNA leading to the cleavage of new target genes that belong to stress responses and cellular metabolic processes. Therefore, the control of AGO1 homeostasis by ubiquitin ligases, plays an important quality control to avoid off-target cleavage.

2022-04-01 | PXD024840 | Pride

The Arabidopsis F-box protein FBW2 degrades AGO1 to avoid spurious loading of illegitimate small RNA [PARE-seq]

Project description:The Arabidopsis F-box protein FBW2 degrades AGO1 to avoid spurious loading of illegitimate small RNA [PARE-seq]

| PRJNA716605 | ENA

The Arabidopsis F-box protein FBW2 degrades AGO1 to avoid spurious loading of illegitimate small RNA

Project description:The Arabidopsis F-box protein FBW2 degrades AGO1 to avoid spurious loading of illegitimate small RNA

| PRJNA716596 | ENA

The Arabidopsis F-box protein FBW2 degrades AGO1 to avoid spurious loading of illegitimate small RNA [ncRNA-Seq]

Project description:The Arabidopsis F-box protein FBW2 degrades AGO1 to avoid spurious loading of illegitimate small RNA [ncRNA-Seq]

| PRJNA716276 | ENA

The Arabidopsis F-box protein FBW2 degrades AGO1 to avoid spurious loading of illegitimate small RNA

Project description:This SuperSeries is composed of the SubSeries listed below.

2022-04-01 | GSE169434 | GEO

Cell cycle dependent regulation and function of Argonaute 1 in plants [PARE]

Project description:The RNA silencing (RNAi) is a biological process, which plays a key role in a wide range of eukaryotes, such as gene expression regulation, transposable elements control and antiviral defense. In human, several studies showed that miRNAs and AGO2 are involved in the regulation of the cell cycle. Indeed, many cell cycle regulators such as cyclins, CDKs, CDK inhibitors, RBR proteins or E2F transcription factors have been identified as miRNA targets. Transcriptional data suggest that these cell cycle miRNAs are differentially accumulated during cell cycle progression and that misregulation of some could lead to proliferative diseases. However, in plants, almost nothing is known about the cell cycle regulation through miRNAs. Unpublished datas from the lab suggest that the Arabidopsis AGO1, which is the most important Argonaute for miRNA-mediated gene regulation, is essential for cell proliferation. In order to investigate the role of AGO1 during cell cycle, we set up a system based on cell cycle synchronization of tobacco BY-2 cell suspensions expressing a GFP-fused AGO1. We sequenced small RNAs from S, G2, M and G1 phases of synchronized cells from total RNA extract and GFP-AGO1 co-purified small RNAs, and investigate whether putative cell cycle specific miRNAs can be highlighted.

2019-06-06 | GSE102343 | GEO

Farnesylated Heat Shock Protein 40 is a component of membrane-bound RISC in Arabidopsis

Project description:ARGONAUTE1 (AGO1) binds directly to small regulatory RNA and is a key effector protein of post-transcriptional gene silencing mediated by microRNA (miRNA) and small interfering RNA (siRNA). The formation of an RNA induced silencing complex (RISC) of AGO1 and small RNA requires the function of the Heat Shock Protein 70/90 chaperone system. Some functions of AGO1 occur in association with endomembranes, in particular the rough endoplasmic reticulum (rER), but proteins interacting with AGO1 in membrane fractions remain unidentified. In this study, we show that the farnesylated Heat Shock Protein 40 homologs, J2 and J3, associate with AGO1 in membrane fractions in a manner that involves protein farnesylation. We also show that three changes in AGO1 function are detectable in mutants in protein farnesylation and J2/J3. First, perturbations of the HSP40/70/90 pathway by mutation of j3, hsp90 and farnesyl transferase affect the amounts of AGO1 associated with membranes. Second, miRNA association with membrane-bound AGO1, and with membrane-bound polysomes is increased in farnesyl transferase and farnesylationdeficient J2/J3 mutants. Third, silencing by non-cell autonomously acting short interfering RNAs (siRNAs) is impaired. These observations highlight the involvement of farnesylated J2/J3 in small RNA-mediated gene regulation, and suggest that the importance of chaperone-AGO1 interaction is not limited to the RISC assembly process.

2018-09-10 | PXD010197 | Pride

The N-terminal extension of Arabidopsis ARGONAUTE 1 is essential for microRNA activities [sRNA-seq]

Project description:We profiled the small RNA of AGO1 NTE truncation mutant lines, and the small RNA associated with AGO1 NTE truncation mutant to study the roles of the NTE region in Arabidopsis AGO1 involved RNA-induced silencing complex (RISC) formation.

2023-02-21 | GSE224480 | GEO

The N-terminal extension of Arabidopsis ARGONAUTE 1 is essential for microRNA activities [ncRNA_seq]

2023-02-21 | GSE212364 | GEO

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