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Heat shock protein 70 together with its co-chaperone CHIP inhibits TNF-α induced apoptosis by promoting proteasomal degradation of apoptosis signal-regulating kinase1

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Abstract

Inducible heat shock protein70 (HSP70) is one of the most important HSPs for maintenance of cell integrity during normal cellular growth as well as pathophysiological conditions. Apoptosis signal-regulating kinase (ASK) 1, a mammalian MAPKKK, activates the JNK and p38 pathways. Here we report a novel function of HSP70 in regulating TNF-α-induced cell apoptosis. Our study demonstrated that HSP70 physically interacted with ASK1 and promoted the ubiquitin-dependent proteasomal degradation of ASK1. CHIP (carboxyl terminus of the HSC70-interacting protein) which acted as a co-chaperone of HSP70 cooperated with HSP70 in regulating ASK1. We also found that TNF-α stimulated HSP70/CHIP/ASK1 association and through cooperating with CHIP, HSP70 inhibits TNF-α-induced cell apoptosis both in over-expression and RNAi conditions. Structural analysis indicated that C-terminal domain of HSP70 was necessary for ASK1 degradation, and N- terminal domain of ASK1 was essential for its binding to HSP70. All these findings indicated that HSP70 and CHIP association is important for HSP70 in interacting with ASK1. Through forming the complex of HSP70/CHIP/ASK1, HSP70 promotes ASK1 proteasomal degradation and prevents TNF-α-induced cell apoptosis.

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Abbreviations

ASK1:

Apoptosis signal-regulating kinase1

HSP70:

Heat shock protein 70

Ub:

Ubiquitin

PARP:

Poly (ADP-ribose) polymerase

CHIP:

Carboxyl terminus of the Hsc70-interacting protein

GAPDH:

Glyceraldehyde phosphate dehydrogenase

References

  1. Ichijo H et al (1997) Induction of apoptosis by ASK1, a mammalian MAPKKK that activates SAPK/JNK and p38 signaling pathways. Science 275:90–94

    Article  CAS  PubMed  Google Scholar 

  2. Tobiume K, Inage T, Takeda K, Enomoto S, Miyazono K, Ichijo H (1997) Molecular cloning and characterization of the mouse apoptosis signal-regulating kinase 1. Biochem Biophys Res Commun 239:905–910

    Article  CAS  PubMed  Google Scholar 

  3. Chang HY, Nishitoh H, Yang X, Ichijo H, Baltimore D (1998) Activation of apoptosis signal-regulating kinase 1 (ASK1) by the adapter protein Daxx. Science 281:1860–1863

    Article  CAS  PubMed  Google Scholar 

  4. Gotoh Y, Cooper JA (1998) Reactive oxygen species- and dimerization-induced activation of apoptosis signal-regulating kinase 1 in tumor necrosis factor-a signal transduction. J Biol Chem 273:17477–17482

    Article  CAS  PubMed  Google Scholar 

  5. Nishitoh H, Saitoh M, Mochida Y, Takeda K, Nakano H, Rothe M, Miyazono K, Ichijo H (1998) ASK1 is essential for JNK/SAPK activation by TRAF2. Mol Cell 2:389–395

    Article  CAS  PubMed  Google Scholar 

  6. 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 (ASK) 1. EMBO J 17:2596–2606

    Article  CAS  PubMed  Google Scholar 

  7. Wang TH, Wang HS, Ichijo H, Giannakakou P, Foster JS, Fojo T, Wimalasena J (1998) Microtubule-interfering agents activate c-Jun N-terminal kinase/stress-activated protein kinase through both Ras and apoptosis signal-regulating kinase pathways. J Biol Chem 273:4928–4936

    Article  CAS  PubMed  Google Scholar 

  8. 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 (TRAF2) requires prior dissociation of the ASK1 inhibitor thioredoxin. Mol Cell Biol 20:2198–2208

    Article  CAS  PubMed  Google Scholar 

  9. Cho SG et al (2001) Glutathione S-Transferase Mu modulates the stress-activated signals by suppressing apoptosis signal-regulating kinase 1. J Biol Chem 276:12749–12755

    Article  CAS  PubMed  Google Scholar 

  10. 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

    Article  CAS  PubMed  Google Scholar 

  11. Zhang R, Luo D, Miao R, Bai L, Ge Q, Sessa WC, Min W (2005) Hsp90-Akt phosphorylates ASK1 and inhibits ASK1-mediated apoptosis. Oncogene 24:3954–3963

    Article  CAS  PubMed  Google Scholar 

  12. Castelli C, Rivoltini L, Rini F, Belli F, Testori A, Maio M, Mazzaferro V, Coppa J, Srivastava PK, Parmiani G (2004) Heat shock proteins: biological functions and clinical application as personalized vaccine for human cancer. Cancer Immunol Immunother 53:227–233

    Article  CAS  PubMed  Google Scholar 

  13. Feder ME, Hofmann GE (1999) Heat-shock proteins, molecular chaperones, and the stress response: evolutionary and ecological physiology. Annu Rev Physiol 61:243–282

    Article  CAS  PubMed  Google Scholar 

  14. Srivastava P (2002) Roles of heat-shock proteins in innate and adaptive immunity. Nat Rev Immunol 2:185–194

    Article  CAS  PubMed  Google Scholar 

  15. Stennicke HR, Ryan CA, Salvesen GS (2002) Reprieval from execution: the molecular basis of caspase inhibition. Trends Biochem Sci 27:94–101

    Article  CAS  PubMed  Google Scholar 

  16. Dudeja V, Mujumdar N, Phillips P, Chugh R, Borja-Cacho D, Dawra RK, Vickers SM, Saluja AK (2009) Heat shock protein 70 inhibits apoptosis in cancer cells through simultaneous and independent mechanisms. Gastroenterology 136:1772–1782

    Article  CAS  PubMed  Google Scholar 

  17. Taleb M, Brandon CS, Lee FS, Harris KC, Dillmann WH, Cunningham LL (2009) Hsp70 inhibits aminoglycoside-induced hearing loss and cochlear hair cell death. Cell Stress Chaperones 14:427–437

    Article  CAS  PubMed  Google Scholar 

  18. Mao H, Li Z, Zhou Y, Li Z, Zhuang S, An X, Zhang B, Chen W, Nie J, Wang Z, Borkan SC, Wang Y, Yu X (2008) HSP72 attenuates renal tubular cell apoptosis and interstitial fibrosis in obstructive nephropathy. Am J Physiol Renal Physiol 295(1):F202–F214

    Article  CAS  PubMed  Google Scholar 

  19. Bienemann AS, Lee YB, Howarth J, Uney JB (2008) Hsp70 suppresses apoptosis in sympathetic neurones by preventing the activation of c-Jun. J Neurochem 104:271–278

    CAS  PubMed  Google Scholar 

  20. Amici C, Rossi A, Satoro MG (1994) Selective inhibition of virus protein synthesis by prostaglandin A: a translational block associated with Hsp70 synthesis. J Virol 68:6890–6899

    CAS  PubMed  Google Scholar 

  21. Mestril R, Chi SH, Sayen MR, O’Reilly K, Dillmann WH (1994) Expression of inducible stress protein70 in rat heart myogenic cells confers protection against stimulated ischemia-induced injury. J Clin Invest 93:759–767

    Article  CAS  PubMed  Google Scholar 

  22. Polla BS, Bachelet M, Elia G, Santoro MG (1998) Stress proteins in inflammation. Ann N Y Acad Sci 851:75–85

    Article  CAS  PubMed  Google Scholar 

  23. Chen H, Wu Y, Zhang Y, Jin L, Luo L, Xue B, Lu C, Zhang X, Yin Z (2006) Hsp70 inhibits lipopolysaccharide-induced NF-κB activation by interacting with TRAF6 and inhibiting its ubiquitination. FEBS Lett 580:3145–3152

    Article  CAS  PubMed  Google Scholar 

  24. Takayama S, Reed JC, Homma S (2003) Heat-shock proteins as regulators of apoptosis. Oncogene 22:9041–9047

    Article  CAS  PubMed  Google Scholar 

  25. Höhfeld J, Cyr DM, Patterson C (2001) From the cradle to the grave: molecular chaperones that may choose between folding and degradation. EMBO Rep 2:885–890

    Article  PubMed  Google Scholar 

  26. Alberti S, Esser C, Höhfeld J (2003) BAG-1–a nucleotide exchange factor of Hsc70 with multiple cellular functions. Cell Stress Chaperones 8:225–231

    Article  PubMed  Google Scholar 

  27. Fan CY, Lee S, Cyr DM (2003) Mechanisms for regulation of Hsp70 function by Hsp40. Cell Stress Chaperones 8:309–316

    Article  CAS  PubMed  Google Scholar 

  28. Odunuga OO, Longshaw VM, Blatch GL (2004) Hop: more than an Hsp70/Hsp90 adaptor protein. BioEssays 26:1058–1068

    Article  CAS  PubMed  Google Scholar 

  29. Ballinger CA, Connell P, Wu Y, Hu Z, Thompson LJ, Yin LY, Patterson C (1999) Identification of CHIP, a novel tetratricopeptide repeat-containing protein that interacts with heat shock proteins and negatively regulates chaperone functions. Mol Cell Biol 19:4535–4545

    CAS  PubMed  Google Scholar 

  30. Jiang J, Ballinger CA, Wu Y, Dai Q, Cyr DM, Hohfeld J, Patterson C (2001) CHIP is a U-box-dependent E3 ubiquitin ligase: identification of Hsc70 as a target for ubiquitylation. J Biol Chem 276:42938–42944

    Article  CAS  PubMed  Google Scholar 

  31. Cardozo CP, Michaud C, Ost MC, Fliss AE, Yang E, Patterson C, Hall SJ, Caplan AJ (2003) C-terminal Hsp-interacting protein slows androgen receptor synthesis and reduces its rate of degradation. Arch Biochem Biophys 410:134–140

    Article  CAS  PubMed  Google Scholar 

  32. Connell P, Ballinger CA, Jiang J, Wu Y, Thompson LJ, HÖhfeld J, Patterson C (2001) The co-chaperone CHIP regulates protein triage decisions mediated by heat-shock proteins. Nat Cell Biol 3:93–96

    Article  CAS  PubMed  Google Scholar 

  33. Hwang JR, Zhang C, Patterson C (2005) C-terminus of heat shock protein 70-interacting protein facilitates degradation of apoptosis signal-regulating kinase 1 and inhibits apoptosis signal-regulating kinase 1-dependent apoptosis. Cell Stress Chaperones 10:147–156

    Article  CAS  PubMed  Google Scholar 

  34. Dewey WC, Freeman ML (1980) Rationale for use of hyperthermia cancer therapy. Ann NY Acad Sci 335:372–378

    Article  CAS  PubMed  Google Scholar 

  35. Beere HM (2004) The stress of dying: the role of heat shock proteins in the regulation of apoptosis. J Cell Sci 117:2641–2651

    Article  CAS  PubMed  Google Scholar 

  36. Nishitoh H, Matsuzawa A, Tobiume K, Saegusa K, Takeda K, Inoue K, Hori S, Kakizuka A, Ichijo H (2002) ASK1 is essential for endoplasmic reticulum stress-induced neuronal cell death triggered by expanded polyglutamine repeats. Genes Dev 16:1345–1355

    Article  CAS  PubMed  Google Scholar 

  37. Glickman MH, Ciechanover A (2002) The ubiquitin-proteasome proteo-lytic pathway: destruction for the sake of construction. Physiol Rev 82:373–428

    CAS  PubMed  Google Scholar 

  38. McDonough H, Patterson C (2003) CHIP: a link between the chaperone and proteasome systems. Cell Stress Chaperones 8:303–308

    Article  CAS  PubMed  Google Scholar 

  39. Jäättelä M, Wissing D, Kokholm K, Kallunki T, Egeblad M (1998) Hsp70 exerts its anti-apoptotic function downstream of caspase-3 -like proteases. EMBO J 17:6124–6134

    Article  PubMed  Google Scholar 

  40. Slee EA, Adrain C, Martin SJ (2001) Executioner caspase-3, -6, and -7 perform distinct, non-redundant roles during the demolition phase of apoptosis. J Biol Chem 276:7320–7326

    Article  CAS  PubMed  Google Scholar 

  41. Koh DW, Ted MD, Valina LD (2005) Mediation of cell death by poly (ADP-ribose) polymerase-1. Pharmacol Res 52:5–14

    Article  CAS  PubMed  Google Scholar 

  42. Garrido C, Schmitt E, Cande C, Vahsen N, Parcellier A, Kroemer G (2003) HSP27 and HSP70: potentially oncogenic apoptosis inhibitors. Cell Cycle 2:579–584

    CAS  PubMed  Google Scholar 

  43. Bukau B, Horwich AL (1998) The Hsp70 and Hsp60 chaperone machines. Cell 92:351–366

    Article  CAS  PubMed  Google Scholar 

  44. Demand J, Alberti S, Patterson C, HÖhfeld J (2001) Cooperation of a ubiquitin domain protein and an E3 ubiquitin ligase during chaperone/proteasome coupling. Curr Biol 11:1569–1577

    Article  CAS  PubMed  Google Scholar 

  45. McDonough H, Patterson C (2003) CHIP: a link between the chaperone and proteasome systems. Cell Stress Chaperones 8:303–308

    Article  CAS  PubMed  Google Scholar 

  46. Shimura H, Schwartz D, Gygi SP, Kosik KS (2004) CHIP-Hsc70 complex ubiquitinates phosphorylated tau and enhances cell survival. J Biol Chem 279:4869–4876

    Article  CAS  PubMed  Google Scholar 

  47. Girnita L, Shenoy SK, Sehat B, Vasilcanu R, Girnita A, Lefkowitz RJ, Larsson O (2005) β-Arrestin is crucial for ubiquitination and down-regulation of the insulin-like growth factor-1 receptor by acting as adaptor for the MDM2 E3 ligase. J Biol Chem 280:24412–24419

    Article  CAS  PubMed  Google Scholar 

  48. Zhang R, Al-Lamki R, Bai L, Streb JW, Miano JM, Bradley J, Min W (2004) Thioredoxin-2 inhibits mitochondria-located ASK1-mediated apoptosis in a JNK-independent manner. Circ Res 94:1483–1491

    Article  CAS  PubMed  Google Scholar 

  49. Takeda K, Matsuzawa A, Nishitoh H, Ichijo H (2003) Roles of MAPKKK ASK1 in stress-induced cell death. Cell Struct Funct 28:23–29

    Article  CAS  PubMed  Google Scholar 

  50. Beere HM, Wolf BB, Cain K, Mosser DD, Mahboubi A, Kuwana T, Tailor P, Morimoto RI, Cohen GM, Green DR (2000) Heat-shock protein 70 inhibits apoptosis by preventing recruitment of procaspase-9 to the Apaf-1 apoptosome. Nat Cell Biol 2:469–475

    Article  CAS  PubMed  Google Scholar 

  51. Mosser DD, Caron AW, Bourget L, Meriin AB, Sherman MY, Morimoto RI, Massie B (2000) The chaperone function of hsp70 is required for protection against stress-induced apoptosis. Mol Cell Biol 20:7146–7159

    Article  CAS  PubMed  Google Scholar 

  52. Saleh A, Srinivasula SM, Balkir L, Robbins PD, Alnemri ES (2000) Negative regulation of the Apaf-1 apoptosome by Hsp70. Nat Cell Biol 2:476–483

    Article  CAS  PubMed  Google Scholar 

  53. Ravagnan L, Gurbuxani S, Susin SA, Maisse C, Daugas E, Zamzami N, Mak T, Jäättelä M, Penninger JM, Garrido C, Kroemer G (2001) Heat-shock protein 70 antagonizes apoptosis-inducing factor. Nat Cell Biol 3:839–843

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We particularly thank professor Dr. Ze’ev Ronai and Chen Wang for providing partial constructs used in this study. This work was supported by grants from National Nature Foundation of China (No. 30770842 and No. 30771979) and Jiangsu Major Nature Science Foundation of High Education (No. 07KJA18026). The Specialized Research Fund for the Doctoral Program of Higher Education (SRFDP), the Ministry of Education of China (2007104SBJ0152).

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Correspondence to Lan Luo or Zhimin Yin.

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Gao, Y., Han, C., Huang, H. et al. Heat shock protein 70 together with its co-chaperone CHIP inhibits TNF-α induced apoptosis by promoting proteasomal degradation of apoptosis signal-regulating kinase1. Apoptosis 15, 822–833 (2010). https://doi.org/10.1007/s10495-010-0495-7

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