No abstract

Caspase-2 appears to be activated by the PIDDosome, but the role of PIDD in this pathway remains controversial. Ando et al. demonstrate that caspase-2 can be activated by two distinct complexes: one in the nucleolus that comprises PIDD and the nucleolar phosphoprotein NPM1, and one in the cytoplasm that is PIDD independent. Therefore, the nucleolus is a novel site for caspase-2 activation that appears to be essential for caspase-2 function.

Drug-based strategies to overcome tumour resistance to radiotherapy (R-RT) remain limited by the single-agent toxicity of traditional radiosensitizers (e.g., platinums) and a lack of targeted alternatives. In a screen for compounds that restore radiosensitivity in p53 mutant zebrafish while tolerated in non-irradiated wild-type animals, we identified the benzimidazole anthelmintic, oxfendazole. Surprisingly, oxfendazole acts via inhibition of IRAK1, a kinase otherwise involved in Interleukin-1 and Toll-like receptor (IL-1R/TLR) immune responses. IRAK1 drives R-RT in a pathway involving IRAK4 and TRAF6 but not the IL-1R/TLR—IRAK adaptor MyD88. Rather than stimulating NF-κB, radiation-activated IRAK1 acts to prevent apoptosis mediated by the PIDDosome complex (PIDD/RAIDD/caspase-2). Countering this pathway with IRAK1 inhibitors suppresses R-RT in tumour models derived from cancers in which TP53 mutations predict R-RT. Lastly, IRAK1 inhibitors synergize with inhibitors of PIN1, a prolyl isomerase essential for IRAK1 activation in response to pathogens and, as shown here, ionizing radiation. These data identify an IRAK1 radiation-response pathway as a rational chemo-RT target.

Summary The PIDDosome—PIDD-RAIDD-caspase-2 complex—is a proapoptotic caspase-activation platform of elusive significance. DNA damage can initiate complex assembly via ATM phosphorylation of the PIDD death domain (DD), which enables RAIDD recruitment to PIDD. In contrast, the mechanisms limiting PIDDosome formation have remained unclear. We identify the mitotic checkpoint factor, BubR1, as a direct PIDDosome inhibitor, acting in a noncanonical role independent of Mad2. Following its phosphorylation by ATM at DNA breaks, ‘primed’ PIDD relocates to kinetochores via a direct interaction with BubR1. BubR1 binds the PIDD DD, competes with RAIDD recruitment, and negates PIDDosome-mediated apoptosis after ionizing radiation. The PIDDosome thus sequentially integrates DNA damage and mitotic checkpoint signals to decide cell fate in response to genotoxic stress. We further show that by sequestering PIDD at the kinetochore, BubR1 acts to delay PIDDosome formation until the next cycle, defining a new mechanism by which cells evade apoptosis during mitosis.