Site-specific DICER and DROSHA RNA products control the DNA-damage response

Sofia Francia, Flavia Michelini, Alka Saxena, Dave Tang, Michiel de Hoon, Viviana Anelli, Marina Mione, Piero Carninci, Fabrizio d'Adda di Fagagna
2012 Nature  
Non-coding RNAs (ncRNAs) are involved in an increasing number of cellular events 1 . Some ncRNAs are processed by DICER and DROSHA ribonucleases to give rise to small doublestranded RNAs involved in RNA interference (RNAi) 2 . The DNA-damage response (DDR) is a signaling pathway that originates from the DNA lesion and arrests cell proliferation 3 . So far, DICER or DROSHA RNA products have not been reported to control DDR activation. Here we show that DICER and DROSHA, but not downstream
more » ... s of the RNAi pathway, are necessary to activate DDR upon oncogene-induced genotoxic stress and exogenous DNA damage, as studied also by DDR foci formation in mammalian cells and zebrafish and by checkpoint assays. DDR foci are sensitive to RNase A treatment, and DICER-and DROSHA-dependent RNA products are required to restore DDR foci in treated cells. Through RNA deep sequencing and studies of DDR activation at an inducible unique DNA double-strand break (DSB), we demonstrate that DDR foci formation requires site-specific DICER-and DROSHA-dependent small RNAs, named DDRNAs, which act in a MRE11-RAD50-NBS1 (MRN) complex-dependent manner. Chemically synthesized or in vitro-generated by DICER cleavage, DDRNAs are sufficient to restore DDR in RNase A-treated cells, also in the absence of other cellular RNAs. Our results describe an unanticipated direct role of a novel class of ncRNAs in the control of DDR activation at sites of DNA damage. § Corresponding author: IFOM Foundation -The FIRC Institute of Molecular Oncology Foundation via Adamello 16, Author Contributions A.S., D.T. and P.C. planned, generated and analysed the genomics data as in S20 a-e, S21, S22b, S23. M. d. H. performed statistical analysis of the genomics data. A.S., P.C. also edited the manuscript. M.M. and V.A. generated data in S14 and S15. F.M. generated data shown in 2b; 3d, e; 4b; S14 d, f; S2 b, e; S3 e; S4b; S5 f, g; S6 b-d; S7 d; S9; S13 d-f; S17 f, g; S18 a,b; S19; S20 g,h; S22a and generated RNA for deep sequencing; contributed to: S16a; S5 d, e; S11 c, d and edited the manuscript. S.F.
doi:10.1038/nature11179 pmid:22722852 pmcid:PMC3442236 fatcat:gel5q5l6tneyjavbua7lf2tcby