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Ago–TNRC6 triggers microRNA-mediated decay by promoting two deadenylation steps | Nature Structural & Molecular Biology
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miRNAs can repress transcripts through decay. Mammalian miRNA-mediated deadenylation is now shown to involve both the Pan2–Pan3 and the Ccr4–Caf1 deadenylases. Such deadenylation can be triggered by tethered Ago or TNRC6 and is followed by decapping of the reporter. MicroRNAs (miRNAs) silence the expression of their mRNA targets mainly by promoting mRNA decay. The mechanism, kinetics and participating enzymes for miRNA-mediated decay in mammalian cells remain largely unclear. Combining the approaches of transcriptional pulsing, RNA tethering, overexpression of dominant-negative mutants, and siRNA-mediated gene knockdown, we show that let-7 miRNA-induced silencing complexes (miRISCs), which contain the proteins Argonaute (Ago) and TNRC6 (also known as GW182), trigger very rapid mRNA decay by inducing accelerated biphasic deadenylation mediated by Pan2–Pan3 and Ccr4–Caf1 deadenylase complexes followed by Dcp1–Dcp2 complex–directed decapping in mammalian cells. When tethered to mRNAs, all four human Ago proteins and TNRC6C are each able to recapitulate the two deadenylation steps. Two conserved human Ago2 phenylalanines (Phe470 and Phe505) are critical for recruiting TNRC6 to promote deadenylation. These findings indicate that promotion of biphasic deadenylation to trigger mRNA decay is an intrinsic property of miRISCs.
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nature portfolio permissions reprints privacy policy advertising social media author information authors nature 431 nature 433 nature 455 nature dcp1–dcp2 complex–directed decapping multiple gw-rich regions mirna-mediated gene silencing microrna-mediated gene silencing sirna-mediated gene knockdown ann-bin shyu author correspondence mirna-mediated translational repression microrna-mediated translational repression early-response-gene mrnas multiple argonaute-binding sites mammalian cell-free system springerlink instant access affect pabp-dependent deadenylation deadenylation steps chyi-ying p-body component gw182 permissions ccr4–caf1 deadenylase complexes personal data mirna-mediated decay au-rich elements molecular biology mammalian p-bodies mirna-mediated repression data protection analyzed data microrna-directed cleavage c-terminal half dcp2 decapping complexes mammalian mrna turnover protein synthesis induced human ago2 binds privacy microrna-dependent localization cold spring harbor purine-rich motifs c-terminal domains c-terminal domain rapid mrna decay cold spring harb
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- Decoding ARE-mediated decay: is microRNA part of the equation?
- How do microRNAs regulate gene expression?
- Mechanisms of post-transcriptional regulation by microRNAs: are the answers in sight?
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