Abstract
Mitochondrial heat shock protein 70 (mtHSP70) is found to play a primary role in cellular defense against physiological stress like exposure to environmental contaminants and helpful in the maintenance of cellular homeostasis by promoting the cell survival. In the present investigation, the environmental-stress-induced increase in mtHSP70 levels along with the quantification of apoptosis signal regulating kinase 1 (ASK1) and thioredoxin (Trx) were measured in the liver mitochondria of grey mullets (Mugil cephalus) collected from the polluted Ennore estuary and the unpolluted Kovalam estuary for a period of 2 years. The results showed elevated lipid peroxide (LPO) and decreased total antioxidant capacity along with the decrease in mitochondrial viability percentage. Mitochondrial HSP70, ASK1, and Trx levels were increased under this stress condition. A 42% increase in LPO levels and 18% decrease in mitochondrial survivality were observed in the polluted-site fish liver mitochondria when compared to the results of unpolluted estuary. We also report that, under observed oxidative stress condition in Ennore fish samples, the ASK1 levels are only moderately elevated (13% increase). This may be due to mitochondrial-HSP70-induced adaptive tolerance signaling for the activation of Trx (22% increase) which suppresses the ASK1 expression thereby promoting the cell survival that leads to the maintenance of the cellular homeostasis.
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Abbreviations
- HSP70:
-
heat shock protein 70
- ASK1:
-
apoptosis signal regulating kinase 1
- Trx:
-
thioredoxin
- LPO:
-
lipid peroxide
- MDA:
-
malondialdehyde
- TAC:
-
total antioxidant capacity
References
Adler V, Yin Z, Fuchs SY, Benezra M, Rosario L, Tew KD, Pincus MR, Sardana M, Henderson CJ, Wolf CR, Davis RJ, Ronai Z (1999) Regulation of JNK signaling by GSTp. EMBO J 18:1321–1334 doi:10.1093/emboj/18.5.1321
Balmer Y, Vensel WH, Tanaka CK, Hurkman WJ, Gelhaye E, Rouhier N, Jacquot JP, Manieri W, Schtermann P, Droux M, Buchanan BB (2004) Thioredoxin links to the regulation of fundamental processes of plant mitochondria. Proc Natl Acad Sci USA 101:2642–2647 doi:10.1073/pnas.0308583101
Basu N, Nakano T, Grau EG, Iwama GK (2001) The effects of cortisol on heat shock protein 70 levels in two fish species. Gen Comp Endocrinol 124:97–105 doi:10.1006/gcen.2001.7688
Bebianno MJ, Geret F, Hoarau P, Serafim MA, Coelho MR, Gnassia-Barelli M, Romeo M (2004) Biomarkers in Ruditapes decussatus: a potential bioindicator species. Biomarkers 9:305–330 doi:10.1080/13547500400017820
Bolliger C, Deloche O, Glick BS, Georgopoulos C, Jeno P, Kronidou N, Horst M, Morishima N, Schatz GA (1994) The mitochondrial homolog of bacterial GrpE interacts with mitochondria Hsp70 is essential for viability. EMBO J 13(8):1998–2006
Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254 doi:10.1016/0003-2697(76)90527-3
Chen Y, Cai J, Murphy TJ, Jones DP (2002) Overexpressed human mitochondrial thioredoxin confers resistance to oxidant induced apoptosis in human osteosarcoma cells. J Biol Chem 277:33242–33248 doi:10.1074/jbc.M202026200
Chinni S, Yallapragda R (2000) Toxicity of copper, cadmium, zinc and lead to Penaeus indicus postlarvae: effects of individual metals. J Environ Biol 21:255–258
Chiueh CC, Andoh T, Chock PB (2005) Induction of thioredoxin and mitochondrial survival proteins mediates preconditioning induced cardioprotection and neuroprotection. Ann N Y Acad Sci 1042:403–418 doi:10.1196/annals.1338.034
Cho SG, Lee YH, Park HS, Ryoo K, Kang KW, Park L (2001) Glutathione S-transferase Mu modulates the stress-activated signals by suppressing apoptosis signal-regulating kinase 1. J Biol Chem 276:12749–12755 doi:10.1074/jbc.M005561200
Erel O (2004) A novel automated direct measurement method for total antioxidant capacity using a new generation, more stable ABTS radical cation. Clin Biochem 37:277–285 doi:10.1016/j.clinbiochem.2003.11.015
Farombi EO, Adelowo OA, Ajimoko YR (2007) Biomarkers of oxidative stress and heavy metal levels as indicators of environmental pollution in African catfish (Clarias gariepinus) from Nigeria Ogun River. Int J Environ Res Public Health 4(2):158–165 doi:10.3390/ijerph2007040011
Fischer W, Bianchi G (1984) FAO identification sheets for fishery purposes. Western Indian Ocean. FAO, Rome
Guo S, Wharton W, Moseley P, Shi H (2007) Heat shock protein 70 regulates cellular redox status by modulating glutathione related enzyme activities. Cell Stress Chaperones 12(3):245–254 doi:10.1379/CSC-265.1
Gutterman DD (2005) Mitochondria and reactive oxygen species: an evolution in function. Circ Res 97:302–304 doi:10.1161/01.RES.0000179773.18195.12
Hsieh CC, Papaconstantinou J (2006) ThioredoxinASK1 complex levels regulate ROS-mediated p38 MAPK pathway activity in livers of aged and long-lived snell dwarf mice. FASEB J 20:259–268 doi:10.1096/fj.05-4376com
Johnson D, Lardy H (1967) Isolation of mitochondria. Methods Enzymol 10:94–96 doi:10.1016/0076-6879(67)10018-9
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680 doi:10.1038/227680a0
Leppa S, Pirkkala L, Chow SC, Eriksson JE, Sistonen L (1997) Thioredoxin is transcriptionally induced upon activation of heat shock factor 2*. J Biol Chem 272:30400–30404 doi:10.1074/jbc.272.48.30400
Lill R, Kispal G (2000) Maturation of cellular Fe-S proteins: an essential function of mitochondria. Trends Biochem Sci 25:352–356 doi:10.1016/S0968-0004(00)01589-9
Liu H, Nishitoh H, Ichijo H, Kyriakis JM (2000) Activation of apoptosis signal-regulating kinase 1 (ASK1) by tumor necrosis factor receptor-associated factor 2 requires prior dissociation of the ASK1 inhibitor thioredoxin. Mol Cell Biol 20:2198–2208 doi:10.1128/MCB.20.6.2198-2208.2000
Lykkesfeldt J (2001) Determination of malondialdehyde as dithiobarbituric acid adduct in biological samples by HPLC with fluorescence detection: comparison with ultraviolet visible spectrophotometry. Clin Chem 47(9):1725–1727
Morimoto RI, Santoro MG (1998) Stress inducible responses to heat shock proteins: new pharmacological targets for cytoprotection. Nature Biotechnol 16:833–838 doi:10.1038/nbt0998-833
Morita K, Saitoh M, Tobiume K, Matsuura H, Enomoto S, Nishitoh H, Ichijo H (2001) Negative feedback regulation of ASK1 by protein phosphatase 5 (PP5) in response to oxidative stress. EMBO J 20:6028–6036 doi:10.1093/emboj/20.21.6028
Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63 doi:10.1016/0022-1759(83)90303-4
Neupert W (1997) Protein import into mitochondria. Annu Rev Biochem 66:863–917 doi:10.1146/annurev.biochem.66.1.863
Nisoli E, Carruba MC (2006) Nitric oxide and mitochondrial biogenesis. J Cell Sci 119:2855–2862 doi:10.1242/jcs.03062
Nordberg J, Arner ES (2001) Reactive oxygen species, antioxidants and the mammalian Thioredoxin system. Free Radic Biol Med 11:1287–1312 doi:10.1016/S0891-5849(01)00724-9
Padmini E, Kavitha M (2003) Seasonal pollution assessment through comparative hydrobiological studies in Ennore and Kovalam estuaries. Ind Hydrobiol 6:139–144
Padmini E, Sudha D (2004) Environmental impact on gill mitochondrial function in Mugil cephalus. Aquacult 5(1):89–92
Padmini E, Usharani M (2008) Impact of seasonal variation on HSP70 expression quantitated in stressed fish hepatocytes. Comp Biochem Physiol Part B 151:278–285 doi:10.1016/j.cbpb.2008.07.011
Padmini E, Vijaya Geetha B (2007a) Seasonal influences on hydrographic parameters and pollution status of Ennore estuary. J Environ Hydrol 15:paper 15
Padmini E, Vijaya Geetha B (2007b) A comparative seasonal pollution assessment study on estuary with respect to metal accumulation in Mugil cephalus. Oceanol Hydrobiol Stud 35:91–103 doi:10.2478/v10009-007-0031-4
Padmini E, Usha Rani M, Vijaya Geetha B (2008a) Differential HSP90α expression in fish hepatocytes from polluted estuary during summer. Fisheries Sci 74(5):1118–1126 doi:10.1111/j.1444-2906.2008.01631.x
Padmini E, Usha Rani M, Vijaya Geetha B (2008b) Studies on antioxidant status in Mugil cephalus in response to heavy metal pollution at Ennore estuary. Environ Monit Assess. doi:10.1007/s10661-008-0430-3
Padmini E, Vijaya Geetha B, Usha Rani M (2008c) Liver oxidative stress modulation shows seasonal impact in Mugil cephalus. Braz J Med Biol Res 41:951–955
Padmini E, Vijaya Geetha B, Usha Rani M (2009) Pollution induced nitrative stress and heat shock protein 70 overexpression in fish liver mitochondria. Sci Tot Environ 407:1307–1317
Park HS, Cho SG, Kim CK, Hwang HS, Noh KT, Kim MS, Huh SH, Kim MJ, Ryoo K, Kim EK, Kang WJ, Lee JS, Seo JS, Ko YG, Kim S, Choi EJ (2002) Heat shock protein Hsp72 is a negative regulator of apoptosis signal-regulating kinase 1. Mol Cell Biol 22:7721–7730 doi:10.1128/MCB.22.22
Pouis G, Montfort WR (2001) Properties and biological activities of thioredoxins. Annu Rev Pharmacol Toxicol 41:261–295 doi:10.1146/annurev.pharmtox.41.1.443
Ryan JA, Hightower LE (1994) Evaluation of heavy-metal ion toxicity in fish cells using a combined stress protein and cytotoxicity assay. Environ Toxicol Chem 13:1231–1240 doi:10.1897/1552-8618(1994)13[1231:EOHITI]2.0.CO;2
Saitoh M, Nishitoh H, Fujii M, Takeda K, Tobiume K, Sawada Y, Kawabata M, Miyazono K, Ichijo H (1998) Mammalian thioredoxin is a direct inhibitor of apoptosis signal regulating kinase (ASK1). EMBO J 17:2596–2606 doi:10.1093/emboj/17.9.2596
Sakurai K, Cederbaum AI (1998) Oxidative stress and cytotoxicity induced by ferric-nitrilotriacetate in Hepg2 cells that express Cytochrome P450 2E1. Mol Pharmacol 54(6):1024–1035
Salinas AE, Wong MG (1999) Glutathione S-transferases—a review. Curr Med Chem 6:279–309
Sanders BM, Martin LS (1993) Stress proteins as biomarkers of contaminant exposure in archived environmental samples. Sci Tot Environ 139–140:459–470 doi:10.1016/0048-9697(93)90043-6
Schatz G, Dobberstein B (1996) Common principles of protein translocation across membrane. Science 271:1519–1526 doi:10.1126/science.271.5255.1519
Simon MM, Reikerstorfer A, Schwatz A, Luger TA, Jaattela M, Schwatz T (1995) Heat shock protein 70 overexpression affects the response to ultraviolet light in murine fibroblasts. Evidence for increased cell viability and suppression of cytokine release. J Clin Investig 95:926–933 doi:10.1172/JCI117800
Thannickal VJ, Fanburg BL (2000) Reactive oxygen species in cell signaling. Am J Physiol 279:L1005–L1028
Tiano L, Davies I, Craft J, Falcioni G (2004) Analysis of stress induced gene expression in trout red blood cells following tributylin chloride exposure. Fish Physiol Biochem 30:231–240 doi:10.1007/s10695-005-8244-5
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354 doi:10.1073/pnas.76.9.4350
Wickner W, Schekman R (2005) Protein translocation across biological membranes. Science 310:1452–1456 doi:10.1126/science.1113752
Zhao Y, Wang W, Qian L (2007) Hsp70 may protect cardiomyocytes from stress-induced injury by inhibiting Fas-mediated apoptosis. Cell Stress Chaperones 12:83–95 doi:10.1379/CSC-231R.1
Acknowledgement
This study is a part of the DST Project Ref. No. DST: SP/SO/AS-10/2003 funded by the Ministry of Science and Technology, Government of India. The Council of Scientific and Industrial Research is acknowledged with thanks for providing Senior Research Fellowship to B. Vijaya Geetha.
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Padmini, E., Vijaya Geetha, B. Modulation of ASK1 expression during overexpression of Trx and HSP70 in stressed fish liver mitochondria. Cell Stress and Chaperones 14, 459–467 (2009). https://doi.org/10.1007/s12192-009-0099-8
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DOI: https://doi.org/10.1007/s12192-009-0099-8