Letter | Published:

Selective autophagy degrades DICER and AGO2 and regulates miRNA activity

Nature Cell Biology volume 14, pages 13141321 (2012) | Download Citation

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  • A Corrigendum to this article was published on 31 July 2015

This article has been updated

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.

Author information

Author notes

    • Derrick Gibbings
    •  & Serge Mostowy

    These authors contributed equally to this work

Affiliations

  1. Swiss Federal Institute of Technology (ETH-Z), Department of Biology, Zürich 8092, Switzerland

    • Derrick Gibbings
    • , Florence Jay
    •  & Olivier Voinnet
  2. University of Ottawa, Department of Cellular and Molecular Medicine, Ottawa K1H 8M5, Canada

    • Derrick Gibbings
  3. Institut Pasteur, Unité des Interactions Bactéries-Cellules, Département de Biologie Cellulaire et Infection, 75015 Paris, France

    • Serge Mostowy
    •  & Pascale Cossart
  4. Inserm, Unité 604, 75015 Paris, France

    • Serge Mostowy
    •  & Pascale Cossart
  5. INRA, USC2020, 75015 Paris, France

    • Serge Mostowy
    •  & Pascale Cossart
  6. Section of Microbiology, MRC Centre for Molecular Bacteriology and Infection, Imperial College London, Armstrong Road, London SW7 2AZ, UK

    • Serge Mostowy
  7. Institut de Génétique et de Biologie Moléculaire et Cellulaire, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 7104, Institut National de la Santé et de la Recherche Médicale Unité 964, 1 rue Laurent Fries, 67404 Illkirch, France

    • Yannick Schwab
  8. Institut de Biologie Moléculaire des Plantes (IBMP), 12 rue du Général Zimmer, 67084 Strasbourg Cedex, France

    • Olivier Voinnet

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Contributions

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.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Derrick Gibbings or Serge Mostowy or Olivier Voinnet.

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DOI

https://doi.org/10.1038/ncb2611

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