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O-GlcNAc modification of eIF4GI acts as a translational switch in heat shock response

Abstract

Heat shock response (HSR) is an ancient signaling pathway leading to thermoprotection of nearly all living organisms. Emerging evidence suggests that intracellular O-linked β-N-acetylglucosamine (O-GlcNAc) serves as a molecular ‘thermometer’ by reporting ambient temperature fluctuations. Whether and how O-GlcNAc modification regulates HSR remains unclear. Here we report that, upon heat shock stress, the key translation initiation factor eIF4GI undergoes dynamic O-GlcNAcylation at the N-terminal region. Without O-GlcNAc modification, the preferential translation of stress mRNAs is impaired. Unexpectedly, stress mRNAs are entrapped within stress granules (SGs) that are no longer dissolved during stress recovery. Mechanistically, we show that stress-induced eIF4GI O-GlcNAcylation repels poly(A)-binding protein 1 and promotes SG disassembly, thereby licensing stress mRNAs for selective translation. Using various eIF4GI mutants created by CRISPR/Cas9, we demonstrate that eIF4GI acts as a translational switch via reversible O-GlcNAcylation. Our study reveals a central mechanism linking heat stress sensing, protein remodeling, SG dynamics and translational reprogramming.

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Fig. 1: Cells lacking OGT exhibit deficient Hsp70 translation.
Fig. 2: eIF4GI undergoes dynamic O-GlcNAcylation in response to heat shock stress.
Fig. 3: eIF4GI O-GlcNAcylation repels PABP1.
Fig. 4: Depletion of O-GlcNAcylation from eIF4GI prevents SG disassembly.
Fig. 5: The N terminus of eIF4GI acts a functional switch via O-GlcNAcylation.
Fig. 6: Rescuing effects of eIF4GI variants in SG dynamics and Hsp70 synthesis.

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Acknowledgements

We thank N. E. Zachara for providing inducible MEF Ogt knockout cell lines. We also thank Qian lab members for discussions. We are grateful to Cornell University Life Sciences Core Laboratory Center for confocal microscope imaging support and mass spectrometry. We thank the Proteomic and MS Facility of Cornell University for providing the mass spectrometry data. The Orbitrap Fusion mass spectrometer was supported by US National Institutes of Health SIG grant 1S10 OD017992-01. This work was supported by grants to S.-B.Q. from the US National Institutes of Health (R01AG042400 and R01GM1222814) and a HHMI Faculty Scholar grant (55108556).

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X.Z. and S.-B.Q conceived the project and designed the experiments. X.Z. performed the majority of experiments. X.E.S. assisted the experiments of eIF4GI isoforms and different chaperones in OGT knockout cells. S.-B.Q wrote the manuscripts. All authors discussed the results and edited the manuscript.

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Correspondence to Shu-Bing Qian.

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Zhang, X., Shu, X.E. & Qian, SB. O-GlcNAc modification of eIF4GI acts as a translational switch in heat shock response. Nat Chem Biol 14, 909–916 (2018). https://doi.org/10.1038/s41589-018-0120-6

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