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
Subscribe to this journal
Receive 12 print issues and online access
$189.00 per year
only $15.75 per issue
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
Similar content being viewed by others
References
Nouspikel, T. & Hanawalt, P.C. DNA repair in terminally differentiated cells. DNA Repair (Amst.) 1, 59–75 (2002).
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).
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).
Belloni, L. et al. DNp73α protects myogenic cells from apoptosis. Oncogene 25, 3606–3612 (2006).
Yaneva, M. & Jhiang, S. Expression of the Ku protein during cell proliferation. Biochim. Biophys. Acta 1090, 181–187 (1991).
Pillai, R.S., Bhattacharyya, S.N. & Filipowicz, W. Repression of protein synthesis by miRNAs: how many mechanisms? Trends Cell Biol. 17, 118–126 (2007).
Ambros, V. The functions of animal microRNAs. Nature 431, 350–355 (2004).
Bartel, D.P. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116, 281–297 (2004).
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).
Fernandez-Capetillo, O., Lee, A., Nussenzweig, M. & Nussenzweig, A. H2AX: the histone guardian of the genome. DNA Repair (Amst.) 3, 959–967 (2004).
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).
Stucki, M. & Jackson, S.P. gammaH2AX and MDC1: anchoring the DNA-damage-response machinery to broken chromosomes. DNA Repair (Amst.) 5, 534–543 (2006).
Bassing, C.H. et al. Histone H2AX: a dosage-dependent suppressor of oncogenic translocations and tumors. Cell 114, 359–370 (2003).
Celeste, A. et al. H2AX haploinsufficiency modifies genomic stability and tumor susceptibility. Cell 114, 371–383 (2003).
Tzur, G. et al. MicroRNA expression patterns and function in endodermal differentiation of human embryonic stem cells. PLoS ONE 3, e3726 (2008).
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).
Sun, Q. et al. Transforming growth factor-β–regulated miR-24 promotes skeletal muscle differentiation. Nucleic Acids Res. 36, 2690–2699 (2008).
Bartel, D.P. MicroRNAs: target recognition and regulatory functions. Cell 136, 215–233 (2009).
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).
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).
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).
Song, E. et al. Sustained small interfering RNA-mediated human immunodeficiency virus type 1 inhibition in primary macrophages. J. Virol. 77, 7174–7181 (2003).
Barad, O. et al. MicroRNA expression detected by oligonucleotide microarrays: system establishment and expression profiling in human tissues. Genome Res. 14, 2486–2494 (2004).
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).
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
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
Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–6 and Supplementary Tables 1 and 2 (PDF 352 kb)
Rights 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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nsmb.1589
This article is cited by
-
Analyzing the impact of 900 MHz EMF short-term exposure to the expression of 667 miRNAs in human peripheral blood cells
Scientific Reports (2021)
-
Loss of miR-24-3p promotes epithelial cell apoptosis and impairs the recovery from intestinal inflammation
Cell Death & Disease (2021)
-
The miR-183/ItgA3 axis is a key regulator of prosensory area during early inner ear development
Cell Death & Differentiation (2017)
-
A role of human RNase P subunits, Rpp29 and Rpp21, in homology directed-repair of double-strand breaks
Scientific Reports (2017)
-
The expression of miRNAs is associated with tumour genome instability and predicts the outcome of ovarian cancer patients treated with platinum agents
Scientific Reports (2017)