Abstract
Multiciliated cells lining the surface of some vertebrate epithelia are essential for various physiological processes, such as airway cleansing1,2,3. However, the mechanisms governing motile cilia biosynthesis remain poorly elucidated. We identify miR-449 microRNAs as evolutionarily conserved key regulators of vertebrate multiciliogenesis. In human airway epithelium and Xenopus laevis embryonic epidermis, miR-449 microRNAs strongly accumulated in multiciliated cells. In both models, we show that miR-449 microRNAs promote centriole multiplication and multiciliogenesis by directly repressing the Delta/Notch pathway. We established Notch1 and its ligand Delta-like 1(DLL1) as miR-449 bona fide targets. Human DLL1 and NOTCH1 protein levels were lower in multiciliated cells than in surrounding cells, decreased after miR-449 overexpression and increased after miR-449 inhibition. In frog, miR-449 silencing led to increased Dll1 expression. Consistently, overexpression of Dll1 mRNA lacking miR-449 target sites repressed multiciliogenesis, whereas both Dll1 and Notch1 knockdown rescued multiciliogenesis in miR-449-deficient cells. Antisense-mediated protection of miR-449-binding sites of endogenous human Notch1 or frog Dll1 strongly repressed multiciliogenesis. Our results unravel a conserved mechanism whereby Notch signalling must undergo miR-449-mediated inhibition to permit differentiation of ciliated cell progenitors.
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Change history
14 September 2011
In the version of this Letter initially published online and in print, an article by Lizé et al. (Cell Cycle 9, 4579–4583; 2010), which reports that miR 449 microRNAs accumulate during mucociliary differentiation of human airway epithelia, was inadvertently omitted from the references list. On pages 1–2, the following text has replaced the previous text: “miR 449a, miR 449b and miR 449c (collectively named miR 449), constitute by far the most strongly induced microRNAs during epithelium differentiation in both species. Although representing less than 0.01% of all microRNA sequences in proliferating HAECs, miR 449 accounted for more than 8% of the microRNA reads in differentiated HAECs (Fig. 1a and Supplementary Fig. S1c,d; see also ref. 13).” The omitted reference has now been added to the reference list: 13. Lizé, M., Herr, C., Klimke, A., Bals, R. & Dobbelstein, M. MicroRNA 449a levels increase by several orders of magnitude during mucociliary differentiation of airway epithelia. Cell Cycle 9, 4579–4583 (2010). References 13–40 have been changed to 14–41, respectively.
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Acknowledgements
This work was supported by CNRS, INSERM, Région Champagne-Ardenne, Région PACA, CG06 and by grants from ANR, Vaincre la Mucoviscidose, ARC and INCa.
We thank V. Magnone, G. Rios, S. Fourré, K. LeBrigand and J. Maurizio, from the IBISA Functional Genomics Platform, S Antipolis, for help with transcriptome analyses and bioinformatics, F. Brau and J. Cazareth, for cellular imaging, V. Thomé for in situ hybridization experiments, F. Aguila for artwork and B. Mari for discussions.
This work is the object of a CNRS patent N°09/03723.
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B.M. led the project. P. Barbry, the Principal Investigator (IPMC), initiated and managed the entire project. L.K. is the Principal Investigator (IBDML) of the Xenopus section. B.M., L.K. and P. Barbry planned experiments, analysed and interpreted data and wrote the paper. B.M. and B. Chevalier carried out cell culture, cellular and molecular biology and cellular imaging in human, C.C., B.N-R. and T.J. carried out in situ hybridization experiments, cell culture and cell sorting on human tissues, G.L., M.C. and L.K. carried out Xenopus experiments, K.R-S. contributed to bioinformatics analyses, L-E.Z. and R.W. carried out HTS experiments, R.W. helped with manuscript correction, B. Cardinaud helped with molecular cloning, C.M. carried out Affymetrix transcriptome experiments and L.G-C. helped with PCR experiments. P. Birembaut provided critical discussion.
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Marcet, B., Chevalier, B., Luxardi, G. et al. Control of vertebrate multiciliogenesis by miR-449 through direct repression of the Delta/Notch pathway. Nat Cell Biol 13, 693–699 (2011). https://doi.org/10.1038/ncb2241
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DOI: https://doi.org/10.1038/ncb2241
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