miRNA repression involves GW182-mediated recruitment of CCR4–NOT through conserved W-containing motifs

Chekulaeva, Marina; Mathys, Hansruedi; Zipprich, Jakob T.; Attig, Jan; Čolić, Marija; Parker, Roy; Filipowicz, Witold

doi:10.1038/nsmb.2166

Cited by 326 publications

(475 citation statements)

References 36 publications

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“…Alternatively, deadenylation may happen independently of the initial translational block but at a slower rate. The stimulated deadenylation, catalysed by PAN2-PAN3 and CCR4-NOT complexes [12] that are directly recruited by the miRISC [3][4][5], may potentiate the effect on translational inhibition and lead to decay of target mRNAs (step 3) through the recruitment of decapping machinery [12,28]. This model, similar to another proposed recently [6], involves three distinct and successive major steps, and thus implies the possibility of reversibility or stalling at the transition from one step to the next.…”

Section: Translational Inhibition Precedes Poly(a) Tail Shorteningmentioning

confidence: 61%

“…Analyses were also performed under deadenylation-impaired conditions, either by simultaneous knockdown of CNOT1 (protein responsible for the recruitment of the CCR4-NOT deadenylation complex by miRISC [3][4][5]), and the deadenylases CNOT7 and CNOT8, or overexpression of dominant-negative mutants of PAN2 and CNOT7, which were shown to efficiently block miRNA-mediated deadenylation of reporters [12]. Neither of the treatments affected the initial B30% repression of the hmga2-WT reporter at 1-h postinduction.…”

Section: Translational Inhibition Precedes Poly(a) Tail Shorteningmentioning

confidence: 99%

“…Bound to AGO, miRNAs serve as guides bringing miRISC to target mRNAs by hybridizing, generally through imperfect base pairing, with their 3 0 -untranslated regions (3 0 UTRs) [1,2]. GW182 proteins (named TNRC6 in vertebrates), acting downstream of AGOs, recruit the CCR4-NOT deadenylation complex, resulting in the downregulation of mRNA function [3][4][5]. It is widely agreed that miRNAs induce the deadenylation and decay of target mRNAs.…”

Section: Introductionmentioning

confidence: 99%

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Kinetic analysis reveals successive steps leading to miRNA‐mediated silencing in mammalian cells

2012

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MicroRNAs (miRNAs) regulate most cellular functions, acting by posttranscriptionally repressing numerous eukaryotic mRNAs. They lead to translational repression, deadenylation and degradation of their target mRNAs. Yet, the relative contributions of these effects are controversial and little is known about the sequence of events occurring during the miRNA-induced response. Using stable human cell lines expressing inducible reporters, we found that translational repression is the dominant effect of miRNAs on newly synthesized targets. This step is followed by mRNA deadenylation and decay, which is the dominant effect at steady state. Our findings have important implications for understanding the mechanism of silencing and reconcile seemingly contradictory data.

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Section: Translational Inhibition Precedes Poly(a) Tail Shorteningmentioning

confidence: 61%

Section: Translational Inhibition Precedes Poly(a) Tail Shorteningmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Kinetic analysis reveals successive steps leading to miRNA‐mediated silencing in mammalian cells

2012

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“…This model is based on the following observations: First, the interaction of GW182 proteins with the CCR4–NOT complex is not only required for degradation but also required for translational repression of miRNA reporters (Braun et al , 2011; Chekulaeva et al , 2011; Fabian et al , 2011; Huntzinger et al , 2013; Zekri et al , 2013; Chen et al , 2014; Mathys et al , 2014). Second, like miRISC, the CCR4–NOT complex represses translation in the absence of deadenylation (Cooke et al , 2010; Braun et al , 2011; Chekulaeva et al , 2011; Bawankar et al , 2013; Zekri et al , 2013). Translational repression by the CCR4–NOT complex can, at least in part, be explained by a direct interaction between the NOT1 subunit and the DEAD‐box ATPase DDX6 (also known as RCK).…”

Section: Introductionmentioning

confidence: 99%

mi RISC and the CCR 4– NOT complex silence mRNA targets independently of 43S ribosomal scanning

et al. 2016

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AbstractmiRNAs associate with Argonaute (AGO) proteins to silence the expression of mRNA targets by inhibiting translation and promoting deadenylation, decapping, and mRNA degradation. A current model for silencing suggests that AGOs mediate these effects through the sequential recruitment of GW182 proteins, the CCR4–NOT deadenylase complex and the translational repressor and decapping activator DDX6. An alternative model posits that AGOs repress translation by interfering with eIF4A function during 43S ribosomal scanning and that this mechanism is independent of GW182 and the CCR4–NOT complex in Drosophila melanogaster. Here, we show that miRNAs, AGOs, GW182, the CCR4–NOT complex, and DDX6/Me31B repress and degrade polyadenylated mRNA targets that are translated via scanning‐independent mechanisms in both human and Dm cells. This and additional observations indicate a common mechanism used by these proteins and miRNAs to mediate silencing. This mechanism does not require eIF4A function during ribosomal scanning.

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“…GW182-Ago interactions promote target degradation via the sequestration of stabilizing poly(A)-binding proteins [100,101]. Moreover, GW182 proteins interact with the CNOT9 subunit of the CCR4-NOT complex via W-motifs (distinct from GW-motifs), thus recruiting deadenylase activity that catalyzes removal of the poly(A) tail and consequently promotes mRNA degradation [96][97][98] (Fig. 2b).…”

Section: Hhmi Author Manuscriptmentioning

confidence: 99%

From guide to target: molecular insights into eukaryotic RNA-interference machinery

Ipsaro

Joshua‐Tor

2015

Nat Struct Mol Biol

218

144

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Since its relatively recent discovery, RNA interference (RNAi) has emerged as a potent, specific, and ubiquitous means of gene regulation. Through a number of pathways that are conserved from yeast to humans, small non-coding RNAs direct molecular machinery to silence gene expression. In this review, we focus on mechanisms and structures that govern RNA silencing in higher organisms. In addition to highlighting recent advances, parallels and differences between RNAi pathways are discussed. Together, the studies reviewed herein reveal the versatility and programmability of RNA-induced Silencing Complexes (RISCs) and emphasize the importance of both upstream biogenesis and downstream silencing factors. Discovery and Biological Perspectives of RNA interferenceRNAi was first described by Fire and Mello in the 1990s when, in an attempt to use antisense RNA to down-regulate gene expression, they observed that double-stranded RNA (dsRNA) was more potent than sense or anti-sense RNA alone [1]. This seminal work boasted robust and specific gene knock down in addition to coining the term "RNA interference" (RNAi). Shortly beforehand, the discovery of individual regulatory RNAs in C. elegans hinted that small, non-coding RNAs might be a pervasive means of gene regulation in higher organisms [2,3]. In the following decade, with the mechanistic insight of Fire and Mello's work in hand, this idea was confirmed and it is now accepted that wellover 1,000 small RNAs are encoded in the human genome that may regulate over 60% of our genes [4,5]. It also became apparent that several seemingly disconnected phenomena are variations of RNAi-type pathways including co-suppression in plants, DNA elimination in Tetrahymena, and quelling in Neurospora [6]. The prevalence of RNAi is impressive and its importance is underscored by the fact that RNAi dysfunction is associated with numerous diseases and disorders including neurological maladies, cancers, and infertility.Shortly after the initial description of RNAi phenomenology, a burst of genetic, biochemical, biophysical, and bioinformatic efforts laid a strong foundation for identifying the molecular pathways, players, and parameters that govern silencing via RNAi. Work from Reprints and permissions information is available online at http://www.nature.com/reprints/index.html. HHMI Author ManuscriptHHMI Author Manuscript HHMI Author Manuscript numerous groups defined the molecular apparatus of RNAi as RISC (RNA-induced Silencing Complex), a ribonucleoprotein complex minimally comprised of a small singlestranded RNA (~20-31 nucleotides) and an Argonaute protein which serves as the effector molecule (reviewed in [7]). In this configuration, the loaded "guide" RNA acts as a specificity determinant that directs Argonaute and any other associated machinery to the target. The guide-loaded Argonaute platform underlies every example in the expansive array of known RNAi pathways in eukaryotes regardless of the source of the guide (structured loci, transposons, viral, etc.), the machinery ...

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miRNA repression involves GW182-mediated recruitment of CCR4–NOT through conserved W-containing motifs

Cited by 326 publications

References 36 publications

Kinetic analysis reveals successive steps leading to miRNA‐mediated silencing in mammalian cells

Kinetic analysis reveals successive steps leading to miRNA‐mediated silencing in mammalian cells

mi RISC and the CCR 4– NOT complex silence mRNA targets independently of 43S ribosomal scanning

From guide to target: molecular insights into eukaryotic RNA-interference machinery

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