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Senescence is an endogenous trigger for microRNA-directed transcriptional gene silencing in human cells

Nature Cell Biology volume 14, pages 266–275 (2012)Cite this article

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Abstract

Cellular senescence is a tumour-suppressor mechanism that is triggered by cancer-initiating or promoting events in mammalian cells. The molecular underpinnings for this stable arrest involve transcriptional repression of proliferation-promoting genes regulated by the retinoblastoma (RB1)/E2F repressor complex. Here, we demonstrate that AGO2, RB1 and microRNAs (miRNAs), as exemplified here by let-7, physically and functionally interact to repress RB1/E2F-target genes in senescence, a process that we call senescence-associated transcriptional gene silencing (SA-TGS). Herein, AGO2 acts as the effector protein for let-7-directed implementation of silent-state chromatin modifications at target promoters, and inhibition of the let-7/AGO2 effector complex perturbs the timely execution of senescence. Thus, we identify cellular senescence as the an endogenous signal of miRNA/AGO2-mediated TGS in human cells. Our results suggest that miRNA/AGO2-mediated SA-TGS may contribute to tumour suppression by stably repressing proliferation-promoting genes in premalignant cancer cells.

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Figure 1: Identification of AGO-bound E2F-target genes and heterochromatin-bound miRNAs.
Figure 2: AGO2 accumulates in the nucleus of senescent cells and is recruited to promoters of repressed E2F-target genes.
Figure 3: AGO2 cooperates with RB1 to regulate E2F-target gene expression.
Figure 4: Depletion of AGO2 delays senescence arrest in W38 fibroblasts.
Figure 5: Overexpression of AGO2 induces proliferative arrest with features of premature senescence.
Figure 6: AGO2 and let-7f cooperate to induce TGS of E2F-target promoters.
Figure 7: Inhibition of let-7f perturbs timely execution of senescence and SA-TGS.
Figure 8: Model for AGO2 and miRNA function in SA-TGS of E2F-target genes.

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Acknowledgements

We would like to thank A. Verdel and M. Yaniv for discussions and critical reading of the manuscript. We are grateful to P. Adams for providing the anti-macroH2A antibody to U.H. and N. Mirani for technical help with histopathology as well as S. Volinia and C. Croce for miRNA profiling and J. Doudement (GenomeQuest, France) for bioinformatics analysis. This work was supported by grants from Ligue Nationale Contre le Cancer (Equipe labellisée), Association for International Cancer Research, Agence Nationale de la Recherche, Association pour la Recherche sur le Cancer (ARC), OdysseyRe and the New Jersey Commission on Cancer Research 09-1124-CCR-EO to U.H. O.B. is a CNRS (Centre National de la Recherche Scientifique) fellow, A.D. Institut National de la Santé et de la Recherche Médicale (INSERM)/Institut Pasteur and M.B. was supported by ARC.

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Author notes

  1. Anne Dejean and Oliver Bischof: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Cell Biology and Infection, Institut Pasteur, Nuclear Organisation and Oncogenesis Unit, F-75015 Paris, France

    Moussa Benhamed, Anne Dejean & Oliver Bischof

  2. INSERM, U993, F-75015 Paris, France

    Moussa Benhamed, Anne Dejean & Oliver Bischof

  3. New Jersey Medical School–University Hospital Cancer Center and Department of Microbiology and Molecular Genetics, UMDNJ-New Jersey Medical School, Newark, New Jersey 07103, USA

    Utz Herbig

  4. Department of Functional Genomics and Cancer, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), BP F-10142-67404 Illkirch Cedex, France

    Tao Ye

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Contributions

O.B. and M.B. conceived the project. M.B. carried out experiments. O.B., M.B. and A.D. analysed the data. U.H. carried out immunohistochemical staining on nevi and melanomas. T.Y. carried out bioinformatic analysis. O.B. and A.D. wrote the manuscript.

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Correspondence to Anne Dejean or Oliver Bischof.

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Benhamed, M., Herbig, U., Ye, T. et al. Senescence is an endogenous trigger for microRNA-directed transcriptional gene silencing in human cells. Nat Cell Biol 14, 266–275 (2012). https://doi.org/10.1038/ncb2443

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