An interdomain helix in IRE1α mediates the conformational change required for the sensor's activation

The unfolded protein response plays an evolutionarily conserved role in homeostasis, and its dysregulation often leads to human disease, including diabetes and cancer. IRE1 alpha is a major transducer that conveys endoplasmic reticulum stress via biochemical signals, yet major gaps persist in our understanding of how the detection of stress is converted to one of several molecular outcomes. It is known that, upon sensing unfolded proteins via its endoplasmic reticulum luminal domain, IRE1 alpha dimerizes and then oligomerizes (often visualized as clustering). Once assembled, the kinase domain trans-autophosphorylates a neighboring IRE1 alpha, inducing a conformational change that activates the RNase effector domain. However, the full details of how the signal is transmitted are not known. Here, we describe a previously unrecognized role for helix aK, located between the kinase and RNase domains of IRE1 alpha, in conveying this critical conformational change. Using constructs containing mutations within this interdomain helix, we show that distinct substitutions affect oligomerization, kinase activity, and the RNase activity of IRE1 alpha differentially. Furthermore, using both biochemical and computational methods, we found that different residues at position 827 specify distinct conformations at distal sites of the protein, such as in the RNase domain. Of importance, an RNase-inactive mutant, L827P, can still dimerize with wildtype monomers, but this mutation inactivates the wildtype molecule and renders leukemic cells more susceptible to stress. We surmise that helix aK is a conduit for the activation of IRE1 alpha in response to stress.

Automated summary

The unfolded protein response is crucial for maintaining homeostasis and its dysregulation can lead to diseases like diabetes and cancer. IRE1 alpha is a key transducer in detecting endoplasmic reticulum stress, but gaps remain in understanding how this stress detection leads to molecular outcomes. Helix aK in IRE1 alpha plays a vital role in transmitting critical conformational changes in response to stress, affecting oligomerization, kinase activity, and RNase activity differently. Specific residues in position 827 of IRE1 alpha specify distinct conformations and mutations can impact its function, highlighting the importance of aK helix in the activation of IRE1 alpha.