Abstract
MicroRNAs (miRNAs) form a class of short RNAs (∼ 21 nucleotides) that post-transcriptionally regulate partially complementary messenger RNAs. Each miRNA may target tens to hundreds of transcripts to control key biological processes. Although the biochemical reactions underpinning miRNA biogenesis and activity are relatively well defined1,2 and the importance of their homeostasis is increasingly evident, the processes underlying regulation of the miRNA pathway in vivo are still largely elusive3. Autophagy, a degradative process in which cytoplasmic material is targeted into double-membrane vacuoles, is recognized to critically contribute to cellular homeostasis. Here, we show that the miRNA-processing enzyme, DICER (also known as DICER1), and the main miRNA effector, AGO2 (also known as eukaryotic translation initiation factor 2C, 2 (EIF2C2)), are targeted for degradation as miRNA-free entities by the selective autophagy receptor NDP52 (also known as calcium binding and coiled-coil domain 2 (CALCOCO2)). Autophagy establishes a checkpoint required for continued loading of miRNA into AGO2; accordingly, NDP52 and autophagy are required for homeostasis and activity of the tested miRNAs. Autophagy also engages post-transcriptional regulation of the DICER mRNA, underscoring the importance of fine-tuned regulation of the miRNA pathway. These findings have implications for human diseases linked to misregulated autophagy, DICER- and miRNA-levels, including cancer.
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Change history
10 July 2015
In the version of this Letter originally published, the TUBA immunoblotting panel and the Coomassie-stained panel was used in Figure 5c (in the dataset corresponding to let-7 antagomir treatment, top set of panels), and reused in Figure 5e, without appropriate acknowledgement. The Coomassie-stained gel was vertically flipped in Figure 5e but the alignment of the lanes was maintained. The TUBA immunoblot and Coomassie-stained membranes represent experimental controls. The p62 panel in Figures 2j and 2k (CQ-treated) was also reused without appropriate attribution. In all cases of reuse of blots between panels, the samples were obtained within one representative experiment and processed in parallel. The authors confirm that all instances of vertical splicing of lanes, for example in Figs 1 and 3, were carried out in full compliance with the journal guidelines. All spliced samples were collected and processed in a single experiment. The original publication was missing Supplementary Fig. S3 containing the uncropped scans of the blots; this has now been included online.
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Acknowledgements
The authors would like to thank M. Johnston (ETH-Zurich, Switzerland) for providing HEK293T cells stably transfected with Tet-inducible Flag–AGO2 as well as the protocol for detection of ubiquitylated Flag–AGO2. Financial support was provided by a core grant from ETH-Z to O.V., and the Pasteur Institute to P.C. S.M. is a Wellcome Trust Research Career Development Fellow. The authors thank K. McGourty and D. Li for helpful discussions.
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D.G. conceived the hypothesis. D.G., S.M., F.J. and Y.S. performed and analysed experiments. D.G., S.M. and O.V. designed the overall research. D.G., S.M., P.C. and O.V. wrote the manuscript.
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Gibbings, D., Mostowy, S., Jay, F. et al. Selective autophagy degrades DICER and AGO2 and regulates miRNA activity. Nat Cell Biol 14, 1314–1321 (2012). https://doi.org/10.1038/ncb2611
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DOI: https://doi.org/10.1038/ncb2611
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