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Competition for XPO5 binding between Dicer mRNA, pre-miRNA and viral RNA regulates human Dicer levels

Nature Structural & Molecular Biology volume 18, pages 323–327 (2011)Cite this article

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Abstract

MicroRNAs (miRNAs) are a class of small, noncoding RNAs that function by regulating gene expression post-transcriptionally. Alterations in miRNA expression can strongly influence cellular physiology. Here we demonstrated cross-regulation between two components of the RNA interference (RNAi) machinery in human cells. Inhibition of exportin-5, the karyopherin responsible for pre-miRNA export, downregulated expression of Dicer, the RNase III required for pre-miRNA maturation. This effect was post-transcriptional and resulted from an increased nuclear localization of Dicer mRNA. In vitro assays and cellular RNA immunoprecipitation experiments showed that exportin-5 interacted directly with Dicer mRNA. Titration of exportin-5 by overexpression of either pre-miRNA or the adenoviral VA1 RNA resulted in loss of Dicer mRNA–exportin-5 interaction and reduction of Dicer level. This saturation also occurred during adenoviral infection and enhanced viral replication. Our study reveals an important cross-regulatory mechanism between pre-miRNA or viral small RNAs and Dicer through exportin-5.

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Figure 1: Regulation of Dicer protein level by XPO5.
Figure 2: XPO5 knockdown results in accumulation of Dicer mRNA in the nucleus.
Figure 3: Dicer mRNA specifically interacts with XPO5 in vivo and in vitro.
Figure 4: Overexpression of pre-miRNA or adenoviral VA1 RNA affects Dicer protein levels in cells.
Figure 5: XPO5 inhibition enhances adenovirus replication.

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Acknowledgements

We are grateful to R. Kiernan and members of the Laboratoire de Virologie Moléculaire for helpful discussions and for critically reading the manuscript, and to G. Akusjärvi (Uppsala University, Uppsala, Sweden), B. Cullen (Duke University, Durham, North Carolina, USA) and I. Macara (University of Virginia, Charlottesville, Virginia, USA) for providing reagents. Work in M.B.'s laboratory was supported by Agence Nationale de Recherche sur le SIDA, SIDACTION, Agence Nationale de la Recherche-BLAN-0040, European Research Council (ERC 250333) and the Fondation pour la Recherche Médicale Equipe labéllisée FRM.

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Authors and Affiliations

  1. Centre National de la Recherche Scientifique (CNRS), Institut de Génétique Humaine UPR1142, Laboratoire de Virologie Moléculaire, Montpellier, France

    Yamina Bennasser, Christine Chable-Bessia, Robinson Triboulet & Monsef Benkirane

  2. Institut de Biologie Moléculaire des Plantes (IBMP), CNRS, Université de Strasbourg, Strasbourg, France

    Derrick Gibbings & Olivier Voinnet

  3. Institut Jacques Monod, Université Paris Diderot, CNRS, Paris, France

    Carole Gwizdek & Catherine Dargemont

  4. Institut de Génétique Moléculaire de Montpellier, Universités de Montpellier I & II, Montpellier, France

    Eric J Kremer

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Contributions

Y.B. and M.B. planned and supervised the project and wrote the paper. Y.B. designed and performed most of the experiments with the help of C.C.-B. R.T. initiated the project. D.G. performed experiment in O.V.'s laboratory. C.G., C.D. and E.J.K. helped perform experiments and provided valuable reagents.

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Correspondence to Yamina Bennasser or Monsef Benkirane.

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The authors declare no competing financial interests.

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Bennasser, Y., Chable-Bessia, C., Triboulet, R. et al. Competition for XPO5 binding between Dicer mRNA, pre-miRNA and viral RNA regulates human Dicer levels. Nat Struct Mol Biol 18, 323–327 (2011). https://doi.org/10.1038/nsmb.1987

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