Abstract
Dicer initiates RNA interference by generating small RNAs involved in various silencing pathways. Dicer participates in centromeric silencing, but its role in the epigenetic regulation of other chromatin domains has not been explored. Here we show that Dicer1 deficiency in Mus musculus leads to decreased DNA methylation, concomitant with increased telomere recombination and telomere elongation. These DNA-methylation defects correlate with decreased expression of Dnmt1, Dnmt3a and Dnmt3b DNA methyltransferases (Dnmts), and methylation levels can be recovered by their overexpression. We identify the retinoblastoma-like 2 protein (Rbl2) as responsible for decreased Dnmt expression in Dicer1-null cells, suggesting the existence of Dicer-dependent small RNAs that target Rbl2. We identify the miR-290 cluster as being downregulated in Dicer1-deficient cells and show that it silences Rbl2, thereby controlling Dnmt expression. These results identify a pathway by which miR-290 directly regulates Rbl2-dependent Dnmt expression, indirectly affecting telomere-length homeostasis.
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Change history
18 June 2008
In the version of this article initially published, the GEO accession number for the array data was not provided. It is GSE11229. The error has been corrected in the HTML and PDF versions of the article.
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Acknowledgements
We are indebted to O. Dominguez and D. Pisano from the Genomics and Bioinformatics Units of the Spanish National Cancer Centre (CNIO), respectively, for help with bisulfite sequencing analysis and various microarray and genome-sequence analyses. R.B. is a staff investigator of the CNIO, S.G. is a Fondo de Investigaciones Sanitarias (FIS) senior scientist. P.M. is a 'Ramon y Cajal' senior scientist. I.J. is a student of the Gulbenkian Ph.D. Program in Biomedicine, supported by FCT/MCT (Portugal). M.A.B.'s laboratory is funded by the MCyT (SAF2005-00277, GEN2001-4856-C13-08), by the Regional Government of Madrid (GR/SAL/0597/2004), European Union (TELOSENS FIGH-CT-2002-00217, INTACT LSHC-CT-2003-506803, ZINCAGE FOOD-CT-2003-506850, RISC-RAD FI6R-CT-2003-508842, MOL CANCER MED LSHC-CT-2004-502943) and the Josef Steiner Cancer Research Award, 2003.
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R.B. performed experiments shown in all manuscript figures as well as designed the experiments together with M.A.B. and S. Gonzalo. S. Gonzalo performed part of the experiments shown in Figure 3c,d and Figure 5. I.J. performed some of the experiments shown in Figure 3a,b. P.M. performed the experiments shown in Figure 3e,f. S. Gonzalez and M.S. generated the miRNA data shown in Figure 7a. S.S. contributed to initial characterization of Dicer1-null telomeric chromatin. E.M. and G.H. provided the Dicer1-null cells and helped with the writing of the manuscript. T.A. and S.M. provided the Dicer1-null skin samples. T.C. and E.L. provided the Dnmt-deficient cells and the Dnmt constructs. P.K. and M.A.B. prepared the manuscript figures. M.A.B., R.B. and S. Gonzalo wrote the paper.
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Benetti, R., Gonzalo, S., Jaco, I. et al. A mammalian microRNA cluster controls DNA methylation and telomere recombination via Rbl2-dependent regulation of DNA methyltransferases. Nat Struct Mol Biol 15, 268–279 (2008). https://doi.org/10.1038/nsmb.1399
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DOI: https://doi.org/10.1038/nsmb.1399