Activation of Gcn2 in Response to Different Stresses

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2017 Aug 4

PMID 28771613

Citations 44

Authors

Silje Anda

Robert Zach

Beata Grallert

Affiliations

Soon will be listed here.

Abstract

All organisms have evolved pathways to respond to different forms of cellular stress. The Gcn2 kinase is best known as a regulator of translation initiation in response to starvation for amino acids. Work in budding yeast has showed that the molecular mechanism of GCN2 activation involves the binding of uncharged tRNAs, which results in a conformational change and GCN2 activation. This pathway requires GCN1, which ensures delivery of the uncharged tRNA onto GCN2. However, Gcn2 is activated by a number of other stresses which do not obviously involve accumulation of uncharged tRNAs, raising the question how Gcn2 is activated under these conditions. Here we investigate the requirement for ongoing translation and tRNA binding for Gcn2 activation after different stresses in fission yeast. We find that mutating the tRNA-binding site on Gcn2 or deleting Gcn1 abolishes Gcn2 activation under all the investigated conditions. These results suggest that tRNA binding to Gcn2 is required for Gcn2 activation not only in response to starvation but also after UV irradiation and oxidative stress.

Citing Articles

An In Vitro System to Study Mammalian GCN2 Regulation.

Masson G, Vinciauskaite V Methods Mol Biol. 2025; 2882:221-234.

PMID: 39992512 DOI: 10.1007/978-1-0716-4284-9_11.

Circadian clock control of interactions between eIF2α kinase CPC-3 and GCN1 with ribosomes regulates rhythmic translation initiation.

Preh E, Ramirez M, Mohan S, Guy C, Bell-Pedersen D Proc Natl Acad Sci U S A. 2025; 122(6):e2411916122.

PMID: 39903114 PMC: 11831163. DOI: 10.1073/pnas.2411916122.

General control nonderepressible 2 (GCN2) as a therapeutic target in age-related diseases.

Altintas O, MacArthur M Front Aging. 2024; 5:1447370.

PMID: 39319345 PMC: 11420162. DOI: 10.3389/fragi.2024.1447370.

Host WD repeat-containing protein 5 inhibits protein kinase R-mediated integrated stress response during measles virus infection.

BenDavid E, Yang C, Zhou Y, Pfaller C, Samuel C, Ma D J Virol. 2024; 98(9):e0102024.

PMID: 39194235 PMC: 11406981. DOI: 10.1128/jvi.01020-24.

Emerging Role of GCN1 in Disease and Homeostasis.

Tatara Y, Kasai S, Kokubu D, Tsujita T, Mimura J, Itoh K Int J Mol Sci. 2024; 25(5).

PMID: 38474243 PMC: 10931611. DOI: 10.3390/ijms25052998.

References

Moir R, Lee J, Haeusler R, Desai N, Engelke D, Willis I . Protein kinase A regulates RNA polymerase III transcription through the nuclear localization of Maf1. Proc Natl Acad Sci U S A. 2006; 103(41):15044-9. PMC: 1622776. DOI: 10.1073/pnas.0607129103. View

Knutsen J, Rodland G, Boe C, Haland T, Sunnerhagen P, Grallert B . Stress-induced inhibition of translation independently of eIF2α phosphorylation. J Cell Sci. 2015; 128(23):4420-7. PMC: 4712817. DOI: 10.1242/jcs.176545. View

Wek S, Zhu S, Wek R . The histidyl-tRNA synthetase-related sequence in the eIF-2 alpha protein kinase GCN2 interacts with tRNA and is required for activation in response to starvation for different amino acids. Mol Cell Biol. 1995; 15(8):4497-506. PMC: 230689. DOI: 10.1128/MCB.15.8.4497. View

Kearsey S, Montgomery S, Labib K, Lindner K . Chromatin binding of the fission yeast replication factor mcm4 occurs during anaphase and requires ORC and cdc18. EMBO J. 2000; 19(7):1681-90. PMC: 310236. DOI: 10.1093/emboj/19.7.1681. View

Dong J, Qiu H, Anderson J, Hinnebusch A . Uncharged tRNA activates GCN2 by displacing the protein kinase moiety from a bipartite tRNA-binding domain. Mol Cell. 2000; 6(2):269-79. DOI: 10.1016/s1097-2765(00)00028-9. View

Grallert B, Boye E . The Gcn2 kinase as a cell cycle regulator. Cell Cycle. 2007; 6(22):2768-72. DOI: 10.4161/cc.6.22.4933. View

Tvegard T, Soltani H, Skjolberg H, Krohn M, Nilssen E, Kearsey S . A novel checkpoint mechanism regulating the G1/S transition. Genes Dev. 2007; 21(6):649-54. PMC: 1820939. DOI: 10.1101/gad.421807. View

Lageix S, Zhang J, Rothenburg S, Hinnebusch A . Interaction between the tRNA-binding and C-terminal domains of Yeast Gcn2 regulates kinase activity in vivo. PLoS Genet. 2015; 11(2):e1004991. PMC: 4335047. DOI: 10.1371/journal.pgen.1004991. View

Udagawa T, Nemoto N, Wilkinson C, Narashimhan J, Jiang L, Watt S . Int6/eIF3e promotes general translation and Atf1 abundance to modulate Sty1 MAPK-dependent stress response in fission yeast. J Biol Chem. 2008; 283(32):22063-75. PMC: 2494926. DOI: 10.1074/jbc.M710017200. View

10.

Caspari T, DAHLEN M, Lindsay H, Hofmann K, Papadimitriou K, Sunnerhagen P . Characterization of Schizosaccharomyces pombe Hus1: a PCNA-related protein that associates with Rad1 and Rad9. Mol Cell Biol. 2000; 20(4):1254-62. PMC: 85258. DOI: 10.1128/MCB.20.4.1254-1262.2000. View

11.

Sattlegger E, Hinnebusch A . Polyribosome binding by GCN1 is required for full activation of eukaryotic translation initiation factor 2{alpha} kinase GCN2 during amino acid starvation. J Biol Chem. 2005; 280(16):16514-21. DOI: 10.1074/jbc.M414566200. View

12.

Sattlegger E, Hinnebusch A . Separate domains in GCN1 for binding protein kinase GCN2 and ribosomes are required for GCN2 activation in amino acid-starved cells. EMBO J. 2000; 19(23):6622-33. PMC: 305848. DOI: 10.1093/emboj/19.23.6622. View

13.

Valbuena N, Rozalen A, Moreno S . Fission yeast TORC1 prevents eIF2α phosphorylation in response to nitrogen and amino acids via Gcn2 kinase. J Cell Sci. 2012; 125(Pt 24):5955-9. DOI: 10.1242/jcs.105395. View

14.

Kirchner S, Ignatova Z . Emerging roles of tRNA in adaptive translation, signalling dynamics and disease. Nat Rev Genet. 2014; 16(2):98-112. DOI: 10.1038/nrg3861. View

15.

Marton M, Vazquez de Aldana C, Qiu H, Chakraburtty K, Hinnebusch A . Evidence that GCN1 and GCN20, translational regulators of GCN4, function on elongating ribosomes in activation of eIF2alpha kinase GCN2. Mol Cell Biol. 1997; 17(8):4474-89. PMC: 232301. DOI: 10.1128/MCB.17.8.4474. View

16.

Marton M, Crouch D, Hinnebusch A . GCN1, a translational activator of GCN4 in Saccharomyces cerevisiae, is required for phosphorylation of eukaryotic translation initiation factor 2 by protein kinase GCN2. Mol Cell Biol. 1993; 13(6):3541-56. PMC: 359824. DOI: 10.1128/mcb.13.6.3541-3556.1993. View

17.

Qiu H, Hu C, Dong J, Hinnebusch A . Mutations that bypass tRNA binding activate the intrinsically defective kinase domain in GCN2. Genes Dev. 2002; 16(10):1271-80. PMC: 186288. DOI: 10.1101/gad.979402. View

18.

Lu W, Laszlo C, Miao Z, Chen H, Wu S . The role of nitric-oxide synthase in the regulation of UVB light-induced phosphorylation of the alpha subunit of eukaryotic initiation factor 2. J Biol Chem. 2009; 284(36):24281-8. PMC: 2782021. DOI: 10.1074/jbc.M109.008821. View

19.

Nemoto N, Udagawa T, Ohira T, Jiang L, Hirota K, Wilkinson C . The roles of stress-activated Sty1 and Gcn2 kinases and of the protooncoprotein homologue Int6/eIF3e in responses to endogenous oxidative stress during histidine starvation. J Mol Biol. 2010; 404(2):183-201. PMC: 4378542. DOI: 10.1016/j.jmb.2010.09.016. View

20.

Ruff M, Krishnaswamy S, Boeglin M, Poterszman A, Mitschler A, Podjarny A . Class II aminoacyl transfer RNA synthetases: crystal structure of yeast aspartyl-tRNA synthetase complexed with tRNA(Asp). Science. 1991; 252(5013):1682-9. DOI: 10.1126/science.2047877. View