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.
This is a preview of subscription content, access via your institution
Access options
Access Nature and 54 other Nature Portfolio journals
Get Nature+, our best-value online-access subscription
$29.99 / 30 days
cancel any time
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on SpringerLink
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1038%2Fs41589-018-0120-6/MediaObjects/41589_2018_120_Fig1_HTML.png)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1038%2Fs41589-018-0120-6/MediaObjects/41589_2018_120_Fig2_HTML.png)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1038%2Fs41589-018-0120-6/MediaObjects/41589_2018_120_Fig3_HTML.png)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1038%2Fs41589-018-0120-6/MediaObjects/41589_2018_120_Fig4_HTML.png)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1038%2Fs41589-018-0120-6/MediaObjects/41589_2018_120_Fig5_HTML.png)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1038%2Fs41589-018-0120-6/MediaObjects/41589_2018_120_Fig6_HTML.png)
Similar content being viewed by others
References
Hardivillé, S. & Hart, G. W. Nutrient regulation of signaling, transcription, and cell physiology by O-GlcNAcylation. Cell Metab. 20, 208–213 (2014).
Bond, M. R. & Hanover, J. A. A little sugar goes a long way: the cell biology of O-GlcNAc. J. Cell Biol. 208, (869–880 (2015).
Yang, X. & Qian, K. Protein O-GlcNAcylation: emerging mechanisms and functions. Nat. Rev. Mol. Cell Biol. 18, 452–465 (2017).
Janetzko, J. & Walker, S. The making of a sweet modification: structure and function of O-GlcNAc transferase. J. Biol. Chem. 289, 34424–34432 (2014).
Keembiyehetty, C. et al. Conditional knock-out reveals a requirement for O-linked N-acetylglucosaminase (O-GlcNAcase) in metabolic homeostasis. J. Biol. Chem. 290, 7097–7113 (2015).
Radermacher, P. T. et al. O-GlcNAc reports ambient temperature and confers heat resistance on ectotherm development. Proc. Natl Acad. Sci. USA 111, 5592–5597 (2014).
Groves, J. A., Lee, A., Yildirir, G. & Zachara, N. E. Dynamic O-GlcNAcylation and its roles in the cellular stress response and homeostasis. Cell Stress Chaperones 18, 535–558 (2013).
Zachara, N. E. et al. Dynamic O-GlcNAc modification of nucleocytoplasmic proteins in response to stress. A survival response of mammalian cells. J. Biol. Chem. 279, 30133–30142 (2004).
Martinez, M. R., Dias, T. B., Natov, P. S. & Zachara, N. E. Stress-induced O-GlcNAcylation: an adaptive process of injured cells. Biochem. Soc. Trans. 45, 237–249 (2017).
Kazemi, Z., Chang, H., Haserodt, S., McKen, C. & Zachara, N. E. O-linked β-N-acetylglucosamine (O-GlcNAc) regulates stress-induced heat shock protein expression in a GSK-3β-dependent manner. J. Biol. Chem. 285, 39096–39107 (2010).
Richter, K., Haslbeck, M. & Buchner, J. The heat shock response: life on the verge of death. Mol. Cell 40, 253–266 (2010).
Bukau, B., Weissman, J. & Horwich, A. Molecular chaperones and protein quality control. Cell 125, 443–451 (2006).
Morimoto, R. I. Regulation of the heat shock transcriptional response: cross talk between a family of heat shock factors, molecular chaperones, and negative regulators. Genes Dev. 12, 3788–3796 (1998).
Anckar, J. & Sistonen, L. Regulation of HSF1 function in the heat stress response: implications in aging and disease. Annu. Rev. Biochem. 80, 1089–1115 (2011).
Zhou, J. et al. Dynamic m6A mRNA methylation directs translational control of heat shock response. Nature 526, 591–594 (2015).
Vera, M. et al. The translation elongation factor eEF1A1 couples transcription to translation during heat shock response. Elife 3, e03164 (2014).
Zander, G. et al. mRNA quality control is bypassed for immediate export of stress-responsive transcripts. Nature 540, 593–596 (2016).
Buchan, J. R. & Parker, R. Eukaryotic stress granules: the ins and outs of translation. Mol. Cell 36, 932–941 (2009).
Anderson, P. & Kedersha, N. Stress granules: the Tao of RNA triage. Trends Biochem. Sci. 33, 141–150 (2008).
Cherkasov, V. et al. Coordination of translational control and protein homeostasis during severe heat stress. Curr. Biol. 23, 2452–2462 (2013).
Protter, D. S. & Parker, R. Principles and properties of stress granules. Trends Cell Biol 26, 668–679 (2016).
Sun, J., Conn, C. S., Han, Y., Yeung, V. & Qian, S. B. PI3K-mTORC1 attenuates stress response by inhibiting cap-independent Hsp70 translation. J. Biol. Chem. 286, 6791–6800 (2011).
Zeidan, Q., Wang, Z., De Maio, A. & Hart, G. W. O-GlcNAc cycling enzymes associate with the translational machinery and modify core ribosomal proteins. Mol. Biol. Cell 21, 1922–1936 (2010).
Ohn, T., Kedersha, N., Hickman, T., Tisdale, S. & Anderson, P. A functional RNAi screen links O-GlcNAc modification of ribosomal proteins to stress granule and processing body assembly. Nat. Cell Biol. 10, 1224–1231 (2008).
Gradi, A. et al. A novel functional human eukaryotic translation initiation factor 4G. Mol. Cell. Biol. 18, 334–342 (1998).
Wang, Z. et al. Extensive crosstalk between O-GlcNAcylation and phosphorylation regulates cytokinesis. Sci. Signal. 3, ra2 (2010).
Sonenberg, N. & Dever, T. E. Eukaryotic translation initiation factors and regulators. Curr. Opin. Struct. Biol. 13, 56–63 (2003).
Howard, A. & Rogers, A. N. Role of translation initiation factor 4G in lifespan regulation and age-related health. Ageing Res. Rev. 13, 115–124 (2014).
Gallie, D. R. The cap and poly(A) tail function synergistically to regulate mRNA translational efficiency. Genes Dev. 5, 2108–2116 (1991).
Preiss, T. & Hentze, M. W. Dual function of the messenger RNA cap structure in poly(A)-tail-promoted translation in yeast. Nature 392, 516–520 (1998).
Yanagiya, A. et al. Requirement of RNA binding of mammalian eukaryotic translation initiation factor 4GI (eIF4GI) for efficient interaction of eIF4E with the mRNA cap. Mol. Cell. Biol. 29, 1661–1669 (2009).
Jain, S. et al. ATPase-modulated stress granules contain a diverse proteome and substructure. Cell 164, 487–498 (2016).
Walters, R. W., Muhlrad, D., Garcia, J. & Parker, R. Differential effects of Ydj1 and Sis1 on Hsp70-mediated clearance of stress granules in Saccharomyces cerevisiae. RNA 21, 1660–1671 (2015).
Coldwell, M. J. & Morley, S. J. Specific isoforms of translation initiation factor 4GI show differences in translational activity. Mol. Cell. Biol. 26, 8448–8460 (2006).
Capotosti, F. et al. O-GlcNAc transferase catalyzes site-specific proteolysis of HCF-1. Cell 144, 376–388 (2011).
Liberman, N. et al. DAP5 associates with eIF2β and eIF4AI to promote Internal Ribosome Entry Site driven translation. Nucleic Acids Res 43, 3764–3775 (2015).
Jackson, R. J., Hellen, C. U. & Pestova, T. V. The mechanism of eukaryotic translation initiation and principles of its regulation. Nat. Rev. Mol. Cell Biol. 11, 113–127 (2010).
Walsh, D. & Mohr, I. Viral subversion of the host protein synthesis machinery. Nat. Rev. Microbiol. 9, 860–875 (2011).
McCormick, C. & Khaperskyy, D. A. Translation inhibition and stress granules in the antiviral immune response. Nat. Rev. Immunol. 17, 647–660 (2017).
Schneider, R. J. & Mohr, I. Translation initiation and viral tricks. Trends Biochem. Sci. 28, 130–136 (2003).
Piron, M., Vende, P., Cohen, J. & Poncet, D. Rotavirus RNA-binding protein NSP3 interacts with eIF4GI and evicts the poly(A) binding protein from eIF4F. EMBO J 17, 5811–5821 (1998).
Riback, J. A. et al. Stress-triggered phase separation is an adaptive, evolutionarily tuned response. Cell 168, 1028–1040.e1019 (2017).
Silvera, D., Formenti, S. C. & Schneider, R. J. Translational control in cancer. Nat. Rev. Cancer 10, 254–266 (2010).
Grabocka, E. & Bar-Sagi, D. Mutant KRAS enhances tumor cell fitness by upregulating stress granules. Cell 167, 1803–1813.e1812 (2016).
Truitt, M. L. & Ruggero, D. New frontiers in translational control of the cancer genome. Nat. Rev. Cancer 16, 288–304 (2016).
Ramirez-Valle, F., Braunstein, S., Zavadil, J., Formenti, S.C. & Schneider, R.J. eIF4GI links nutrient sensing by mTOR to cell proliferation and inhibition of autophagy. J. Cell Biol. 181, 293–307 (2008).
Sanjana, N. E., Shalem, O. & Zhang, F. Improved vectors and genome-wide libraries for CRISPR screening. Nat. Methods 11, 783–784 (2014).
Shalem, O. et al. Genome-scale CRISPR-Cas9 knockout screening in human cells. Science 343, 84–87 (2014).
Thomas, C. J., Cleland, T. P., Zhang, S., Gundberg, C. M. & Vashishth, D. Identification and characterization of glycation adducts on osteocalcin. Anal. Biochem. 525, 46–53 (2017).
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).
Author information
Authors and Affiliations
Contributions
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.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interests.
Additional information
Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–9
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/s41589-018-0120-6
This article is cited by
-
m6A governs length-dependent enrichment of mRNAs in stress granules
Nature Structural & Molecular Biology (2023)
-
Dynamic eIF3a O-GlcNAcylation controls translation reinitiation during nutrient stress
Nature Chemical Biology (2022)
-
Discovery proteomics defines androgen-regulated glycoprotein networks in prostate cancer cells, as well as putative biomarkers of prostatic diseases
Scientific Reports (2021)
-
O-GlcNAcylation: the “stress and nutrition receptor” in cell stress response
Cell Stress and Chaperones (2021)
-
Proteomic profiling and genome-wide mapping of O-GlcNAc chromatin-associated proteins reveal an O-GlcNAc-regulated genotoxic stress response
Nature Communications (2020)