dsDNA Breaks and Genome Editing Trigger Ribosome Remodeling and Translational Shutdown

DNA damage activates a robust transcriptional stress response, but much less is known about its impact on translation. The advent of genome editing via Cas9-induced DNA double-strand breaks has intensified interest in understanding cellular responses to DNA damage. Here we find that DNA double-strand breaks (DSBs) induced by Cas9 and other damaging agents lead to a reduction of core ribosomal proteins, RPS27A and RPL40, and that the loss of these proteins is post-transcriptional and p53-independent. DSBs furthermore leads to the shutdown of translation through phosphorylation of eukaryotic initiation factor 2 alpha, and bypassing or enhancing this response affects genome editing outcomes. The DSB translational response is widespread and precedes the canonical transcriptional response. Our results demonstrate that even a single double-strand break can lead to ribosome remodeling and reduced translational output, and suggest caution in interpreting cellular phenotypes measured immediately after genome editing.