SRP198393 PRJNA542891 GSE131201 ZAKa Recognizes Stalled Ribosomes through Partially Redundant Sensor Domains Impairment of ribosome function activates the MAPKKK ZAK, leading to activation of mitogen-activated protein (MAP) kinases p38 and JNK and inflammatory signaling. The mechanistic basis for activation of this ribotoxic stress response (RSR) re-mains completely obscure. We show that the long isoform of ZAK (ZAKa) directly associates with ribosomes by inserting its flexible C terminus into the ribosomal intersubunit space. Here, ZAKa binds helix14 of 18S ribosomal RNA (rRNA). An adjacent domainin ZAKa also probes the ribosome, and together, these sensor domains are critically required for RSR activation after inhibition of both the E-site, the peptidyl transferase center (PTC), and ribotoxinaction. Finally, we show that ablation of the RSR response leads to organismal phenotypes and decreased lifespan in the nematode Caenorhabditis elegans (C. elegans). Our findings yield mechanistic insight into how cells detect ribotoxic stress and provide experimental in vivo evidence for its physiological importance. Overall design: 2 samples from Zaka pull-down experiments. GSE131201 pubmed 32289254