Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

miR-24–mediated downregulation of H2AX suppresses DNA repair in terminally differentiated blood cells

Abstract

Terminally differentiated cells have a reduced capacity to repair double-stranded breaks, but the molecular mechanism behind this downregulation is unclear. Here we find that miR-24 is upregulated during postmitotic differentiation of hematopoietic cell lines and regulates the histone variant H2AX, a protein that has a key role in the double-stranded break response. We show that the H2AX 3′ untranslated region contains conserved miR-24 binding sites that are indeed regulated by miR-24. During terminal differentiation, both H2AX mRNA and protein levels are substantially reduced by miR-24 upregulation in in vitro differentiated cells; similar diminished levels are found in primary human blood cells. miR-24–mediated suppression of H2AX renders cells hypersensitive to γ-irradiation and genotoxic drugs, a phenotype that is fully rescued by overexpression of miR-24–insensitive H2AX. Therefore, miR-24 upregulation in postreplicative cells reduces H2AX and makes them vulnerable to DNA damage.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Purchase on Springer Link

Instant access to full article PDF

Prices may be subject to local taxes which are calculated during checkout

Figure 1: miR-24 is upregulated during hematopoietic cell differentiation into multiple lineages.
Figure 2: miR-24 downregulates H2AX expression during terminal differentiation.
Figure 3: miR-24 expression impedes DSB repair and induces chromosomal instability of γ-irradiated K562 cells.
Figure 4: Cells overexpressing miR-24 are hypersensitive to DNA damage by cytotoxic drugs.
Figure 5: Antagonizing miR-24 enhances cell resistance to bleomycin.

Similar content being viewed by others

References

  1. Nouspikel, T. & Hanawalt, P.C. DNA repair in terminally differentiated cells. DNA Repair (Amst.) 1, 59–75 (2002).

    Article  CAS  Google Scholar 

  2. Lukas, C. et al. DNA damage-activated kinase Chk2 is independent of proliferation or differentiation yet correlates with tissue biology. Cancer Res. 61, 4990–4993 (2001).

    CAS  PubMed  Google Scholar 

  3. Puri, P.L. & Sartorelli, V. Regulation of muscle regulatory factors by DNA-binding, interacting proteins, and post-transcriptional modifications. J. Cell. Physiol. 185, 155–173 (2000).

    Article  CAS  Google Scholar 

  4. Belloni, L. et al. DNp73α protects myogenic cells from apoptosis. Oncogene 25, 3606–3612 (2006).

    Article  CAS  Google Scholar 

  5. Yaneva, M. & Jhiang, S. Expression of the Ku protein during cell proliferation. Biochim. Biophys. Acta 1090, 181–187 (1991).

    Article  CAS  Google Scholar 

  6. Pillai, R.S., Bhattacharyya, S.N. & Filipowicz, W. Repression of protein synthesis by miRNAs: how many mechanisms? Trends Cell Biol. 17, 118–126 (2007).

    Article  CAS  Google Scholar 

  7. Ambros, V. The functions of animal microRNAs. Nature 431, 350–355 (2004).

    Article  CAS  Google Scholar 

  8. Bartel, D.P. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116, 281–297 (2004).

    Article  CAS  Google Scholar 

  9. Sevignani, C., Calin, G.A., Siracusa, L.D. & Croce, C.M. Mammalian microRNAs: a small world for fine-tuning gene expression. Mamm. Genome 17, 189–202 (2006).

    Article  CAS  Google Scholar 

  10. Fernandez-Capetillo, O., Lee, A., Nussenzweig, M. & Nussenzweig, A. H2AX: the histone guardian of the genome. DNA Repair (Amst.) 3, 959–967 (2004).

    Article  CAS  Google Scholar 

  11. Petrini, J.H. & Stracker, T.H. The cellular response to DNA double-strand breaks: defining the sensors and mediators. Trends Cell Biol. 13, 458–462 (2003).

    Article  CAS  Google Scholar 

  12. Stucki, M. & Jackson, S.P. gammaH2AX and MDC1: anchoring the DNA-damage-response machinery to broken chromosomes. DNA Repair (Amst.) 5, 534–543 (2006).

    Article  CAS  Google Scholar 

  13. Bassing, C.H. et al. Histone H2AX: a dosage-dependent suppressor of oncogenic translocations and tumors. Cell 114, 359–370 (2003).

    Article  CAS  Google Scholar 

  14. Celeste, A. et al. H2AX haploinsufficiency modifies genomic stability and tumor susceptibility. Cell 114, 371–383 (2003).

    Article  CAS  Google Scholar 

  15. Tzur, G. et al. MicroRNA expression patterns and function in endodermal differentiation of human embryonic stem cells. PLoS ONE 3, e3726 (2008).

    Article  Google Scholar 

  16. Neilson, J.R., Zheng, G.X., Burge, C.B. & Sharp, P.A. Dynamic regulation of miRNA expression in ordered stages of cellular development. Genes Dev. 21, 578–589 (2007).

    Article  CAS  Google Scholar 

  17. Sun, Q. et al. Transforming growth factor-β–regulated miR-24 promotes skeletal muscle differentiation. Nucleic Acids Res. 36, 2690–2699 (2008).

    Article  CAS  Google Scholar 

  18. Bartel, D.P. MicroRNAs: target recognition and regulatory functions. Cell 136, 215–233 (2009).

    Article  CAS  Google Scholar 

  19. Mannironi, C., Bonner, W.M. & Hatch, C.L. H2A.X. a histone isoprotein with a conserved C-terminal sequence, is encoded by a novel mRNA with both DNA replication type and polyA 3′ processing signals. Nucleic Acids Res. 17, 9113–9126 (1989).

    Article  CAS  Google Scholar 

  20. Sandberg, R., Neilson, J.R., Sarma, A., Sharp, P.A. & Burge, C.B. Proliferating cells express mRNAs with shortened 3′ untranslated regions and fewer microRNA target sites. Science 320, 1643–1647 (2008).

    Article  CAS  Google Scholar 

  21. Calin, G.A. et al. MicroRNA profiling reveals distinct signatures in B cell chronic lymphocytic leukemias. Proc. Natl. Acad. Sci. USA 101, 11755–11760 (2004).

    Article  CAS  Google Scholar 

  22. Song, E. et al. Sustained small interfering RNA-mediated human immunodeficiency virus type 1 inhibition in primary macrophages. J. Virol. 77, 7174–7181 (2003).

    Article  CAS  Google Scholar 

  23. Barad, O. et al. MicroRNA expression detected by oligonucleotide microarrays: system establishment and expression profiling in human tissues. Genome Res. 14, 2486–2494 (2004).

    Article  CAS  Google Scholar 

  24. Moorhead, P.S., Nowell, P.C., Mellman, W.J., Battips, D.M. & Hungerford, D.A. Chromosome preparations of leukocytes cultured from human peripheral blood. Exp. Cell Res. 20, 613–616 (1960).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported in part by the US National Institutes of Health grant AI070302 and a GSK-IDI Alliance grant (to J.L.) and by a Barr Award (to D.C.). We thank members of the Lieberman and Chowdhury laboratories for useful discussions.

Author information

Authors and Affiliations

Authors

Contributions

Most of the experiments were performed collaboratively by A.L., Y.P. and F.N. Chromosome breakage analysis was done by L.M. Several constructs used in the study were made by D.M.D. Z.B. and E.M. generated and analyzed the miRNA microarray data. J.L. and D.C. wrote the paper and conceived all the experiments with A.L.

Corresponding authors

Correspondence to Judy Lieberman or Dipanjan Chowdhury.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–6 and Supplementary Tables 1 and 2 (PDF 352 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lal, A., Pan, Y., Navarro, F. et al. miR-24–mediated downregulation of H2AX suppresses DNA repair in terminally differentiated blood cells. Nat Struct Mol Biol 16, 492–498 (2009). https://doi.org/10.1038/nsmb.1589

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nsmb.1589

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing