c- Jun -N Terminal Kinase-Mediated Degradation of γ-Glutamylcysteine Ligase Catalytic Subunit Inhibits GSH Recovery After Acetaminophen Treatment: Role in Sustaining JNK Activation and Liver Injury
Restricted accessResearch articleFirst published online June, 2023
c- Jun -N Terminal Kinase-Mediated Degradation of γ-Glutamylcysteine Ligase Catalytic Subunit Inhibits GSH Recovery After Acetaminophen Treatment: Role in Sustaining JNK Activation and Liver Injury
Acetaminophen (APAP) overdose is the most common cause of acute liver failure in the United States. Liver glutathione (GSH) depletion and sustained P-JNK (c-Jun-N-terminal kinase) activation are key modulators in the mechanism leading to hepatic necrosis. GSH depletion is directly related to the consumption of GSH by APAP metabolites N-acetyl-p-benzoquinone imine (NAPQI). We previously noticed that the glutamate–cysteine ligase catalytic subunit (GCLC), the rate-limiting enzyme in GSH synthesis, rapidly decreased at the same time P-JNK increased. Our aims were to determine if JNK was directly responsible for decreased GCLC causing impaired recovery of GSH and if this was an important factor in determining APAP hepatotoxicity.
Results:
Immunoprecipitation of JNK after APAP identified binding to GCLC. Expression of a site-directed mutated canonical JNK docking site in GCLC was resistant to degradation and led to rapid restoration of GSH and inhibited sustained JNK activation. The JNK-resistant GCLC markedly protected against necrosis and alanine aminotransferase (ALT) elevation. The proteolytic loss of GCLC was abrogated by inhibition of the proteasome, ubiquitination, or calpain.
Innovation:
Using mutated-GCLC resistant to JNK-induced degradation, the results allowed us to identify impaired GSH recovery as an important contributor to early progression of APAP toxicity after the metabolism of APAP and initial GSH depletion had occurred.
Conclusion:
Activated JNK interacts directly with GCLC and leads to proteolytic degradation of GCLC. Degradation of GCLC impairs GSH recovery after APAP allowing the continued activation of JNK. Conversely, rapid recovery of GSH inhibits the sustained activation of the mitogen-activated protein (MAP) kinase cascade and dampens APAP toxicity by suppressing the continued activation of JNK. Antioxid. Redox Signal. 38, 1071–1081.
Get full access to this article
View all access options for this article.
References
1.
AdelusiOB, RamachandranA, LemastersJJ, et al.The role of Iron in lipid peroxidation and protein nitration during acetaminophen-induced liver injury in mice. Toxicol Appl Pharmacol, 2022; 445:116043; doi: 10.1016/j.taap.2022.116043
2.
AgarwalR, MacMillan-CrowLA, RaffertyTM, et al.Acetaminophen-induced hepatotoxicity in mice occurs with inhibition of activity and nitration of mitochondrial manganese superoxide dismutase. J Pharmacol Exp Ther, 2011; 337(1):110–116; doi: 10.1124/jpet.110.176321
3.
AkakpoJY, RamachandranA, DuanL, et al.Delayed treatment with 4-methylpyrazole protects against acetaminophen hepatotoxicity in mice by inhibition of c-Jun n-terminal kinase. Toxicol Sci, 2019; 170(1):57–68; doi: 10.1093/toxsci/kfz077
4.
AlbanoE, RundgrenM, HarvisonPJ, et al.Mechanisms of N-acetyl-p-benzoquinone imine cytotoxicity. Mol Pharmacol, 1985; 28(3):306–311.
5.
BogoyevitchMA, KobeB. Uses for JNK: The many and varied substrates of the c-Jun N-terminal kinases. Microbiol Mol Biol Rev, 2006; 70(4):1061–1095; doi: 10.1128/MMBR.00025-06
6.
BunchorntavakulC, ReddyKR. Acetaminophen (APAP or N-acetyl-p-aminophenol) and acute liver failure. Clin Liver Dis, 2018; 22(2):325–346; doi: 10.1016/j.cld.2018.01.007
7.
ChenY, LiuK, ZhangJ, et al.c-Jun NH2-terminal protein kinase phosphorylates the Nrf2-ECH homology 6 domain of nuclear factor erythroid 2-related factor 2 and downregulates cytoprotective genes in acetaminophen-induced liver injury in mice. Hepatology, 2020; 71(5):1787–1801; doi: 10.1002/hep.31116
8.
ClagueMJ, UrbéS. Ubiquitin: Same molecule, different degradation pathways. Cell, 2010; 143(5):682–685. doi: 10.1016/j.cell.2010.11.012
9.
DeLeveLD, KaplowitzN. Glutathione metabolism and its role in hepatotoxicity. Pharmacol Ther, 1991; 52(3):287–305; doi: 10.1016/0163-7258(91)90029-l
10.
GunawanBK, LiuZX, HanD, et al.c-Jun N-terminal kinase plays a major role in murine acetaminophen hepatotoxicity. Gastroenterology, 2006; 131(1):165–178; doi: 10.1053/j.gastro.2006.03.045
11.
HanD, HanawaN, SaberiB, et al.Mechanisms of liver injury. III. Role of glutathione redox status in liver injury. Am J Physiol Gastrointest Liver Physiol, 2006; 291(1):G1–G7. doi: 10.1152/ajpgi.00001.2006
12.
HanawaN, ShinoharaM, SaberiB, et al.Role of JNK translocation to mitochondria leading to inhibition of mitochondria bioenergetics in acetaminophen-induced liver injury. J Biol Chem, 2008; 283(20):13565–13577; doi: 10.1074/jbc.M708916200
13.
JaeschkeH, XieY, McGillMR. Acetaminophen-induced liver injury: From animal models to humans. J Clin Transl Hepatol, 2014; 2(3):153–161; doi: 10.14218/JCTH.2014.00014
14.
JamesLP, McCulloughSS, LampsLW, et al.Effect of N-acetylcysteine on acetaminophen toxicity in mice: Relationship to reactive nitrogen and cytokine formation. Toxicol Sci, 2003; 75(2):458–467; doi: 10.1093/toxsci/kfg181
15.
JanYH, HeckDE, DragomirAC, et al.Acetaminophen reactive intermediates target hepatic thioredoxin reductase. Chem Res Toxicol, 2014; 27(5):882–894; doi: 10.1021/tx5000443
16.
KaplowitzN, AwTY, OokhtensM. The regulation of hepatic glutathione. Annu Rev Pharmacol Toxicol, 1985; 25:715–744. doi: 10.1146/annurev.pa.25.040185.003435
17.
LeeJI, KangJ, StipanukMH. Differential regulation of glutamate-cysteine ligase subunit expression and increased holoenzyme formation in response to cysteine deprivation. Biochem J, 2006; 393(Pt 1):181–190; doi: 10.1042/BJ20051111
18.
LeeKK, ImaizumiN, ChamberlandSR, et al.Targeting mitochondria with methylene blue protects mice against acetaminophen-induced liver injury. Hepatology, 2015; 61(1):326–336; doi: 10.1002/hep.27385
19.
LeeWM. Acetaminophen (APAP) hepatotoxicity—Isn't it time for APAP to go away?. J Hepatol, 2017; 67(6):1324–1331; doi: 10.1016/j.jhep.2017.07.005
20.
LeeWM, KaplowitzN. Alcohol, fasting, and therapeutic dosing of acetaminophen: A perfect storm. Hepatology, 2021; 73(5):1634–1636; doi: 10.1002/hep.31747
21.
LimayePB, ApteUM, ShankarK, et al.Calpain released from dying hepatocytes mediates progression of acute liver injury induced by model hepatotoxicants. Toxicol Appl Pharmacol, 2003; 191(3):211–226; doi: 10.1016/s0041-008x(03)00250-3
22.
LimayePB, BhaveVS, PalkarPS, et al.Upregulation of calpastatin in regenerating and developing rat liver: Role in resistance against hepatotoxicity. Hepatology, 2006; 44(2):379–388; doi: 10.1002/hep.21250
23.
LouvetA, Ntandja WandjiLC, LemaîtreE, et al.Acute liver injury with therapeutic doses of acetaminophen: A prospective study. Hepatology, 2021; 73(5):1945–1955; doi: 10.1002/hep.31678
McGillMR, LebofskyM, NorrisHR, et al.Plasma and liver acetaminophen-protein adduct levels in mice after acetaminophen treatment: Dose-response, mechanisms, and clinical implications. Toxicol Appl Pharmacol, 2013; 269(3):240–249; doi: 10.1016/j.taap.2013.03.026
26.
MehendaleHM, LimayePB. Calpain: A death protein that mediates progression of liver injury. Trends Pharmacol Sci, 2005; 26(5):232–236; doi: 10.1016/j.tips.2005.03.008
27.
NakagawaH, MaedaS, HikibaY, et al.Deletion of apoptosis signal-regulating kinase 1 attenuates acetaminophen-induced liver injury by inhibiting c-Jun N-terminal kinase activation. Gastroenterology, 2008; 135(4):1311–1321; doi: 10.1053/j.gastro.2008.07.006
28.
OnoY, SaidoTC, SorimachiH. Calpain research for drug discovery: Challenges and potential. Nat Rev Drug Discov, 2016; 15(12):854–876; doi: 10.1038/nrd.2016.212
29.
PattersonAD, CarlsonBA, LiF, et al.Disruption of thioredoxin reductase 1 protects mice from acute acetaminophen-induced hepatotoxicity through enhanced NRF2 activity. Chem Res Toxicol, 2013; 26(7):1088–1096; doi: 10.1021/tx4001013
30.
PierceRH, FranklinCC, CampbellJS, et al.Cell culture model for acetaminophen-induced hepatocyte death in vivo. Biochem Pharmacol, 2002; 64(3):413–424; doi: 10.1016/s0006-2952(02)01180-2
31.
RechsteinerM. PEST sequences are signals for rapid intracellular proteolysis. Semin Cell Biol, 1990; 1(6):433–440.
32.
RechsteinerM, RogersSW. PEST sequences and regulation by proteolysis. Trends Biochem Sci, 1996; 21(7):267–271.
33.
RogersS, WellsR, RechsteinerM. Amino acid sequences common to rapidly degraded proteins: The PEST hypothesis. Science, 1986; 234(4774):364–368; doi: 10.1126/science.2876518
34.
RonaiZA. Monoubiquitination in proteasomal degradation. Proc Natl Acad Sci U S A, 2016; 113(32):8894–8896; doi: 10.1073/pnas.1610186113
35.
SaitoC, LemastersJJ, JaeschkeH. c-Jun N-terminal kinase modulates oxidant stress and peroxynitrite formation independent of inducible nitric oxide synthase in acetaminophen hepatotoxicity. Toxicol Appl Pharmacol, 2010a;246(1–2):8–17; doi: 10.1016/j.taap.2010.04.015
36.
SaitoC, ZwingmannC, JaeschkeH. Novel mechanisms of protection against acetaminophen hepatotoxicity in mice by glutathione and N-acetylcysteine. Hepatology, 2010b;51(1):246–254; doi: 10.1002/hep.23267
37.
SharmaM, GadangV, JaeschkeA. Critical role for mixed-lineage kinase 3 in acetaminophen-induced hepatotoxicity. Mol Pharmacol, 2012; 82(5):1001–1007; doi: 10.1124/mol.112.079863
38.
ShinoharaM, YbanezMD, WinS, et al.Silencing glycogen synthase kinase-3beta inhibits acetaminophen hepatotoxicity and attenuates JNK activation and loss of glutamate cysteine ligase and myeloid cell leukemia sequence 1. J Biol Chem, 2010; 285(11):8244–8255; doi: 10.1074/jbc.M109.054999
ThanTA, LouH, JiC, et al.Role of cAMP-responsive element-binding protein (CREB)-regulated transcription coactivator 3 (CRTC3) in the initiation of mitochondrial biogenesis and stress response in liver cells. J Biol Chem, 2011; 286(25):22047–22054; doi: 10.1074/jbc.M111.240481
41.
WinS, MinRW, ChenCQ, et al.Expression of mitochondrial membrane-linked SAB determines severity of sex-dependent acute liver injury. J Clin Invest, 2019; 129(12):5278–5293; doi: 10.1172/JCI128289
42.
WinS, ThanTA, Fernandez-ChecaJC, et al.JNK interaction with SAB mediates ER stress induced inhibition of mitochondrial respiration and cell death. Cell Death Dis, 2014; 5(1):e989; doi: 10.1038/cddis.2013.522
43.
WinS, ThanTA, HanD, et al.c-Jun N-terminal kinase (JNK)-dependent acute liver injury from acetaminophen or tumor necrosis factor (TNF) requires mitochondrial SAB protein expression in mice. J Biol Chem, 2011; 286(40):35071–35078; doi: 10.1074/jbc.M111.276089
44.
WinS, ThanTA, KaplowitzN. The regulation of JNK signaling pathways in cell death through the interplay with mitochondrial SAB and upstream post-translational effects. Int J Mol Sci, 2018a;19(11):3657; doi: 10.3390/ijms19113657
45.
WinS, ThanTA, MinRW, et al.c-Jun N-terminal kinase mediates mouse liver injury through a novel SAB (SH3BP5)-dependent pathway leading to inactivation of intramitochondrial Src. Hepatology, 2016; 63(6):1987–2003; doi: 10.1002/hep.28486
46.
WinS, ThanTA, ZhangJ, et al.New insights into the role and mechanism of c-Jun-N-terminal kinase signaling in the pathobiology of liver diseases. Hepatology, 2018b;67(5):2013–2024; doi: 10.1002/hep.29689
47.
XieY, RamachandranA, BreckenridgeDG, et al.Inhibitor of apoptosis signal-regulating kinase 1 protects against acetaminophen-induced liver injury. Toxicol Appl Pharmacol, 2015; 286(1):1–9; doi: 10.1016/j.taap.2015.03.019
48.
ZhangJ, MinRWM, LeK, et al.The role of MAP2 kinases and p38 kinase in acute murine liver injury models. Cell Death Dis, 2017; 8(6):e2903; doi: 10.1038/cddis.2017.295
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
