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bioRxiv · 10.1101/2025.09.11.675390

Allosteric Disordering of eIF2B Regulates the Integrated Stress Response

Abstract

The ternary complex (TC), composed of translation initiation factor eIF2, GTP, and initiator methionyl tRNA, delivers the first amino acid to the ribosome to initiate protein synthesis. The activity of the decameric eukaryotic initiation factor 2B complex (eIF2B) initiates TC assembly by catalyzing GDP to GTP exchange on eIF2, thereby setting the TC levels in the cell. Stress-induced phosphorylation converts eIF2 from the substrate of the GDP/GTP exchange reaction into an inhibitor (eIF2-P) of eIF2B. This conversion reduces the cells TC levels and induces the widespread reprogramming of translation known as the Integrated Stress Response (ISR). Here, we chart an allosteric axis running through eIF2B, revealing the importance of a protrusive -helix in its {beta}-subunit, the latch-helix, that locks onto the -subunit to induce eIF2B activity. eIF2-P binding unhooks the latch-helix, opening eIF2B, which inhibits its GDP/GTP exchange activity. Distinct viral proteins have convergently evolved to bind to eIF2B and stabilize the latch-helix-bound active state. Using these insights, we generated ISR-ACTivating compounds, ISRACTs, that stabilize eIF2B in its inhibited, unlatched state. Our study thus highlights how state-transitions in eIF2B are regulated via long-range allostery.

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BibTeXRIS

Dalwadi, U., Subramanian, A., Deal, A., Conrad, J., Venkatesh, M., Boone, M., Egea, P., He, L., Jain, N., Lee, J., Liu, Y., Reineke, L., Saito, K., Talledge, N., Toutkoushian, H., Le Vasseur, M., Zappa, F., de Groot, R., Acosta-Alvear, D., Arthur, C., Nunnari, J., Costa-Mattioli, M., Crawford, J., van Kuppeveld, F. J. M., Croll, T., Walter, P., Frost, A.. 2025-09-16. Allosteric Disordering of eIF2B Regulates the Integrated Stress Response. https://doi.org/10.1101/2025.09.11.675390

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