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Structural insights into the human GW182-PABC interaction in microRNA-mediated deadenylation

Nature Structural & Molecular Biology volume 17, pages 238240 (2010) | Download Citation

Abstract

GW182-family proteins are essential for microRNA-mediated translational repression and deadenylation in animal cells. Here we show that a conserved motif in the human GW182 paralog TNRC6C interacts with the C-terminal domain of polyadenylate binding protein 1 (PABC) and present the crystal structure of the complex. Mutations at the complex interface impair mRNA deadenylation in mammalian cell extracts, suggesting that the GW182-PABC interaction contributes to microRNA-mediated gene silencing.

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Acknowledgements

We thank D. King (Howard Hughes Medical Institute Mass Spectrometry Laboratory, Univ. California Berkeley) for peptide synthesis and mass spectrometry, and C. Ralston and J. Holton (Beamlines 8.2.1 and 8.3.1, Advanced Light Source, Lawrence Berkeley National Laboratory) for assistance with X-ray data collection. We are indebted to W. Filipowicz for discussions and to members of the Doudna laboratory for critical reading of the manuscript. M.J. is supported by a Human Frontiers Science Program fellowship. M.R.F. is supported by a Terry Fox Foundation fellowship from the Canadian Cancer Society. This work was funded in part by a Canadian Institutes of Health Research grant to N.S. N.S. is an International Scholar of the Howard Hughes Medical Institute. J.A.D. is a Howard Hughes Medical Institute Investigator.

Author information

Affiliations

  1. Department of Molecular and Cell Biology, University of California, Berkeley, California, USA.

    • Martin Jinek
    • , Scott M Coyle
    •  & Jennifer A Doudna
  2. Department of Biochemistry, McGill University, Montreal, Quebec, Canada.

    • Marc R Fabian
    •  & Nahum Sonenberg
  3. Goodman Cancer Centre, McGill University, Montreal, Quebec, Canada.

    • Marc R Fabian
    •  & Nahum Sonenberg
  4. Howard Hughes Medical Institute, University of California, Berkeley, California, USA.

    • Scott M Coyle
    •  & Jennifer A Doudna
  5. Department of Chemistry, University of California, Berkeley, California, USA.

    • Jennifer A Doudna
  6. Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.

    • Jennifer A Doudna

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Contributions

M.J., M.R.F., N.S. and J.A.D. designed experiments. M.J. performed binding assays, crystallized the TNRC6C–PABC complex and determined its structure. M.R.F. performed in vitro deadenylation assays. S.M.C. assisted with X-ray data collection and structure determination. M.J. and J.A.D. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Jennifer A Doudna.

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    Supplementary Figures 1–6, Supplementary Table 1 and Supplementary Methods

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DOI

https://doi.org/10.1038/nsmb.1768

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