[Show abstract][Hide abstract] ABSTRACT: DNA double-strand breaks (DSBs) are highly cytotoxic lesions and pose a major threat to genome stability if not properly repaired. We and others have previously shown that a class of DSB-induced small RNAs (diRNAs) is produced from sequences around DSB sites. DiRNAs are associated with Argonaute (Ago) proteins and play an important role in DSB repair, though the mechanism through which they act remains unclear. Here, we report that the role of diRNAs in DSB repair is restricted to repair by homologous recombination (HR) and that it specifically relies on the effector protein Ago2 in mammalian cells. Interestingly, we show that Ago2 forms a complex with Rad51 and that the interaction is enhanced in cells treated with ionizing radiation. We demonstrate that Rad51 accumulation at DSB sites and HR repair depend on catalytic activity and small RNA-binding capability of Ago2. In contrast, DSB resection as well as RPA and Mre11 loading is unaffected by Ago2 or Dicer depletion, suggesting that Ago2 very likely functions directly in mediating Rad51 accumulation at DSBs. Taken together, our findings suggest that guided by diRNAs, Ago2 can promote Rad51 recruitment and/or retention at DSBs to facilitate repair by HR.Cell Research advance online publication 25 March 2014; doi:10.1038/cr.2014.36.
Cell Research 03/2014; 24(5). DOI:10.1038/cr.2014.36 · 11.98 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: DNA double-strand break (DSB) is the most deleterious form of DNA damage and poses great threat to genome stability. Eukaryotes have evolved complex mechanisms to repair DSBs through coordinated actions of protein sensors, transducers, and effectors. DSB-induced small RNAs (diRNAs) or Dicer/Drosha-dependent RNAs (DDRNAs) have been recently discovered in plants and vertebrates, adding an unsuspected RNA component into the DSB repair pathway. DiRNAs/DDRNAs control DNA damage response (DDR) activation by affecting DDR foci formation and cell cycle checkpoint enforcement and are required for efficient DSB repair. Here, we summarize the findings of diRNAs/DDRNAs and discuss the possible mechanisms through which they act to facilitate DSB repair.
[Show abstract][Hide abstract] ABSTRACT: Eukaryotes have evolved complex mechanisms to repair DNA double-strand breaks (DSBs) through coordinated actions of protein sensors, transducers, and effectors. Here we show that ∼21-nucleotide small RNAs are produced from the sequences in the vicinity of DSB sites in Arabidopsis and in human cells. We refer to these as diRNAs for DSB-induced small RNAs. In Arabidopsis, the biogenesis of diRNAs requires the PI3 kinase ATR, RNA polymerase IV (Pol IV), and Dicer-like proteins. Mutations in these proteins as well as in Pol V cause significant reduction in DSB repair efficiency. In Arabidopsis, diRNAs are recruited by Argonaute 2 (AGO2) to mediate DSB repair. Knock down of Dicer or Ago2 in human cells reduces DSB repair. Our findings reveal a conserved function for small RNAs in the DSB repair pathway. We propose that diRNAs may function as guide molecules directing chromatin modifications or the recruitment of protein complexes to DSB sites to facilitate repair.