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PMID: 10982831 Published · ppublish English Journal Article

The chaperone function of hsp70 is required for protection against stress-induced apoptosis.

Molecular and cellular biology ·Vol. 20 ·No. 19 ·2000-10-00 ·Pages 7146-59

Mosser DD, Caron AW, Bourget L, Meriin AB, Sherman MY, Morimoto RI, Massie B

Abstract

Cellular stress can trigger a process of self-destruction known as apoptosis. Cells can also respond to stress by adaptive changes that increase their ability to tolerate normally lethal conditions. Expression of the major heat-inducible protein hsp70 protects cells from heat-induced apoptosis. hsp70 has been reported to act in some situations upstream or downstream of caspase activation, and its protective effects have been said to be either dependent on or independent of its ability to inhibit JNK activation. Purified hsp70 has been shown to block procaspase processing in vitro but is unable to inhibit the activity of active caspase 3. Since some aspects of hsp70 function can occur in the absence of its chaperone activity, we examined whether hsp70 lacking its ATPase domain or the C-terminal EEVD sequence that is essential for peptide binding was required for the prevention of apoptosis. We generated stable cell lines with tetracycline-regulated expression of hsp70, hsc70, and chaperone-defective hsp70 mutants lacking the ATPase domain or the C-terminal EEVD sequence or containing AAAA in place of EEVD. Overexpression of hsp70 or hsc70 protected cells from heat shock-induced cell death by preventing the processing of procaspases 9 and 3. This required the chaperone function of hsp70 since hsp70 mutant proteins did not prevent procaspase processing or provide protection from apoptosis. JNK activation was inhibited by both hsp70 and hsc70 and by each of the hsp70 domain mutant proteins. The chaperoning activity of hsp70 is therefore not required for inhibition of JNK activation, and JNK inhibition was not sufficient for the prevention of apoptosis. Release of cytochrome c from mitochondria was inhibited in cells expressing full-length hsp70 but not in cells expressing the protein with ATPase deleted. Together with the recently identified ability of hsp70 to inhibit cytochrome c-mediated procaspase 9 processing in vitro, these data demonstrate that hsp70 can affect the apoptotic pathway at the levels of both cytochrome c release and initiator caspase activation and that the chaperone function of hsp70 is required for these effects.

MeSH Terms
Adaptation, Physiological Adenosine Triphosphatases/chemistry,deficiency,genetics Amino Acid Sequence Amino Acid Substitution Apoptosis/physiology Carrier Proteins/physiology Caspase 3 Caspase 9 Caspases/metabolism Cell Division Cell Line Cytochrome c Group/metabolism Enzyme Activation Enzyme Precursors/metabolism HSC70 Heat-Shock Proteins HSP70 Heat-Shock Proteins/chemistry,deficiency,genetics,physiology Hot Temperature Humans JNK Mitogen-Activated Protein Kinases Mitochondria/enzymology Mitogen-Activated Protein Kinases/antagonists & inhibitors Mutagenesis, Site-Directed Protein Folding Protein Structure, Tertiary Recombinant Fusion Proteins/biosynthesis,chemistry,physiology Signal Transduction Stress, Physiological/metabolism,pathology Structure-Activity Relationship Transfection
Chemicals
Carrier Proteins Cytochrome c Group Enzyme Precursors HSC70 Heat-Shock Proteins HSP70 Heat-Shock Proteins HSPA8 protein, human Recombinant Fusion Proteins JNK Mitogen-Activated Protein Kinases Mitogen-Activated Protein Kinases CASP3 protein, human CASP9 protein, human Caspase 3 Caspase 9 Caspases Adenosine Triphosphatases
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Mosser D D
Biotechnology Research Institute, Montreal, Quebec H4P 2R2, Canada. [email protected]
Caron A W
Bourget L
Meriin A B
Sherman M Y
Morimoto R I
Massie B
References (70)
70 references, click to expand
  1. Bcl-2 inhibits the mitochondrial release of an apoptogenic protease.
    J Exp Med. 1996 Oct 1;184(4):1331-41 PMID: 8879205
  2. Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked.
    Science. 1997 Feb 21;275(5303):1129-32 PMID: 9027314
  3. Requirement for ceramide-initiated SAPK/JNK signalling in stress-induced apoptosis.
    Nature. 1996 Mar 7;380(6569):75-9 PMID: 8598911
  4. The carboxy-terminal domain of Hsc70 provides binding sites for a distinct set of chaperone cofactors.
    Mol Cell Biol. 1998 Apr;18(4):2023-8 PMID: 9528774
  5. Importance of the ATP-binding domain and nucleolar localization domain of HSP72 in the protection of nuclear proteins against heat-induced aggregation.
    Exp Cell Res. 1994 Sep;214(1):279-84 PMID: 8082731
  6. Use of a dicistronic expression cassette encoding the green fluorescent protein for the screening and selection of cells expressing inducible gene products.
    Biotechniques. 1997 Jan;22(1):150-4, 156, 158-61 PMID: 8994662
  7. Heat shock protein hsp70 accelerates the recovery of heat-shocked mammalian cells through its modulation of heat shock transcription factor HSF1.
    Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):2126-30 PMID: 7892235
  8. Signal transduction by the c-Jun N-terminal kinase (JNK)--from inflammation to development.
    Curr Opin Cell Biol. 1998 Apr;10(2):205-19 PMID: 9561845
  9. The DNA-binding activity of the human heat shock transcription factor is regulated in vivo by hsp70.
    Mol Cell Biol. 1993 Sep;13(9):5427-38 PMID: 8355691
  10. Expression of human HSP70 during the synthetic phase of the cell cycle.
    Proc Natl Acad Sci U S A. 1986 Dec;83(24):9517-21 PMID: 3540942
  11. Inhibition of cellular proliferation by the Wilms tumor suppressor WT1 requires association with the inducible chaperone Hsp70.
    Genes Dev. 1998 Apr 15;12(8):1108-20 PMID: 9553041
  12. Reduced thermotolerance in aged cells results from a loss of an hsp72-mediated control of JNK signaling pathway.
    Cell Stress Chaperones. 1998 Dec;3(4):265-71 PMID: 9880239
  13. Protein kinase cascades activated by stress and inflammatory cytokines.
    Bioessays. 1996 Jul;18(7):567-77 PMID: 8757935
  14. Heat-shock protein 70 inhibits apoptosis by preventing recruitment of procaspase-9 to the Apaf-1 apoptosome.
    Nat Cell Biol. 2000 Aug;2(8):469-75 PMID: 10934466
  15. Identification of a regulatory motif in Hsp70 that affects ATPase activity, substrate binding and interaction with HDJ-1.
    EMBO J. 1995 May 15;14(10):2281-92 PMID: 7774586
  16. Heat shock proteins delivered with a virus vector can protect cardiac cells against apoptosis as well as against thermal or hypoxic stress.
    J Mol Cell Cardiol. 1999 Jan;31(1):135-46 PMID: 10072722
  17. The function of HSP72 in suppression of c-Jun N-terminal kinase activation can be dissociated from its role in prevention of protein damage.
    J Biol Chem. 1999 Jul 16;274(29):20223-8 PMID: 10400639
  18. Over-expression of hsp70 confers tumorigenicity to mouse fibrosarcoma cells.
    Int J Cancer. 1995 Mar 3;60(5):689-93 PMID: 7860144
  19. Stress-signalling kinase Sek1 protects thymocytes from apoptosis mediated by CD95 and CD3.
    Nature. 1997 Jan 23;385(6614):350-3 PMID: 9002521
  20. Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis.
    Science. 1995 Nov 24;270(5240):1326-31 PMID: 7481820
  21. Hsp70 exerts its anti-apoptotic function downstream of caspase-3-like proteases.
    EMBO J. 1998 Nov 2;17(21):6124-34 PMID: 9799222
  22. The Hsp70 and Hsp60 chaperone machines.
    Cell. 1998 Feb 6;92(3):351-66 PMID: 9476895
  23. The role of inducible 70-kDa heat shock protein in cell cycle control, differentiation, and apoptotic cell death of the human myeloid leukemic HL-60 cells.
    Cell Immunol. 1998 Jul 10;187(1):1-12 PMID: 9681997
  24. Regulation of the heat shock transcriptional response: cross talk between a family of heat shock factors, molecular chaperones, and negative regulators.
    Genes Dev. 1998 Dec 15;12(24):3788-96 PMID: 9869631
  25. Heat shock proteins increase resistance to apoptosis.
    Exp Cell Res. 1996 Feb 25;223(1):163-70 PMID: 8635489
  26. Major heat shock protein hsp70 protects tumor cells from tumor necrosis factor cytotoxicity.
    EMBO J. 1992 Oct;11(10):3507-12 PMID: 1396553
  27. Mitochondria and apoptosis.
    Science. 1998 Aug 28;281(5381):1309-12 PMID: 9721092
  28. In vivo heat shock protects rat myocardial mitochondria.
    Biochem Biophys Res Commun. 1998 May 29;246(3):836-40 PMID: 9618299
  29. Bcl-2 proteins: regulators of apoptosis or of mitochondrial homeostasis?
    Nat Cell Biol. 1999 Dec;1(8):E209-16 PMID: 10587660
  30. Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: inhibition by overexpression of Bcl-2 and Abl.
    J Exp Med. 1995 Nov 1;182(5):1545-56 PMID: 7595224
  31. Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade.
    Cell. 1997 Nov 14;91(4):479-89 PMID: 9390557
  32. Cytochrome c and dATP-mediated oligomerization of Apaf-1 is a prerequisite for procaspase-9 activation.
    J Biol Chem. 1999 Jun 18;274(25):17941-5 PMID: 10364241
  33. Cell cycle-dependent association of HSP70 with specific cellular proteins.
    J Cell Biol. 1989 Feb;108(2):413-23 PMID: 2645297
  34. Thermal response of rat fibroblasts stably transfected with the human 70-kDa heat shock protein-encoding gene.
    Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1681-5 PMID: 1705702
  35. Oncogenic potential of Hsp72.
    Oncogene. 1999 Jun 17;18(24):3648-51 PMID: 10380887
  36. Molecular chaperones in cellular protein folding.
    Nature. 1996 Jun 13;381(6583):571-9 PMID: 8637592
  37. The release of cytochrome c from mitochondria: a primary site for Bcl-2 regulation of apoptosis.
    Science. 1997 Feb 21;275(5303):1132-6 PMID: 9027315
  38. Heat shock proteins and cell proliferation in human breast cancer biopsy samples.
    Cancer Detect Prev. 1997;21(5):441-51 PMID: 9307847
  39. Role of Hsp70 in regulation of stress-kinase JNK: implications in apoptosis and aging.
    FEBS Lett. 1998 Oct 30;438(1-2):1-4 PMID: 9821948
  40. T cell lymphoma in transgenic mice expressing the human Hsp70 gene.
    Biochem Biophys Res Commun. 1996 Jan 17;218(2):582-7 PMID: 8561799
  41. Expression of heat shock protein 70 blocks thymic differentiation of T cells in transgenic mice.
    Immunology. 1998 Dec;95(4):559-65 PMID: 9893045
  42. The development of thermotolerance protects blowfly flight muscle mitochondrial function from heat damage
    J Exp Biol. 1995;198(Pt 11):2413-21 PMID: 9320335
  43. Role of the human heat shock protein hsp70 in protection against stress-induced apoptosis.
    Mol Cell Biol. 1997 Sep;17(9):5317-27 PMID: 9271409
  44. Transcriptional activation by tetracyclines in mammalian cells.
    Science. 1995 Jun 23;268(5218):1766-9 PMID: 7792603
  45. The stress-activated protein kinase pathway mediates cell death following injury induced by cis-platinum, UV irradiation or heat.
    Curr Biol. 1996 May 1;6(5):606-13 PMID: 8805279
  46. Overexpression of the heat shock protein 70 enhances the TCR/CD3- and Fas/Apo-1/CD95-mediated apoptotic cell death in Jurkat T cells.
    J Immunol. 1997 Jun 15;158(12):5668-75 PMID: 9190915
  47. Heat shock protein 70 inhibits apoptosis downstream of cytochrome c release and upstream of caspase-3 activation.
    J Biol Chem. 2000 Aug 18;275(33):25665-71 PMID: 10806214
  48. Heat shock protects neuronal cells from programmed cell death by apoptosis.
    Neuroscience. 1993 Aug;55(3):621-7 PMID: 8413925
  49. Effects of expressing human Hsp70 and its deletion derivatives on heat killing and on RNA and protein synthesis.
    Exp Cell Res. 1995 Apr;217(2):460-8 PMID: 7535238
  50. Protein-damaging stresses activate c-Jun N-terminal kinase via inhibition of its dephosphorylation: a novel pathway controlled by HSP72.
    Mol Cell Biol. 1999 Apr;19(4):2547-55 PMID: 10082520
  51. ATPase activity of the heat shock protein hsp72 is dispensable for its effects on dephosphorylation of stress kinase JNK and on heat-induced apoptosis.
    FEBS Lett. 1999 Nov 12;461(1-2):73-6 PMID: 10561499
  52. The role of c-Jun N-terminal kinase (JNK) in apoptosis induced by ultraviolet C and gamma radiation. Duration of JNK activation may determine cell death and proliferation.
    J Biol Chem. 1996 Dec 13;271(50):31929-36 PMID: 8943238
  53. Use of a micromanipulator for high-efficiency cloning of cells co-expressing fluorescent proteins.
    Methods Cell Sci. 2000;22(2-3):137-45 PMID: 11264945
  54. Molecular chaperones as HSF1-specific transcriptional repressors.
    Genes Dev. 1998 Mar 1;12(5):654-66 PMID: 9499401
  55. Mitochondria are selective targets for the protective effects of heat shock against oxidative injury.
    Proc Natl Acad Sci U S A. 1996 Jun 25;93(13):6458-63 PMID: 8692837
  56. Hsp70 prevents activation of stress kinases. A novel pathway of cellular thermotolerance.
    J Biol Chem. 1997 Jul 18;272(29):18033-7 PMID: 9218432
  57. The temporal relationship between protein phosphatase, mitochondrial cytochrome c release, and caspase activation in apoptosis.
    Exp Cell Res. 1999 Mar 15;247(2):505-13 PMID: 10066378
  58. The consequences of expressing hsp70 in Drosophila cells at normal temperatures.
    Genes Dev. 1992 Aug;6(8):1402-13 PMID: 1644286
  59. Heat shock resistance conferred by expression of the human HSP27 gene in rodent cells.
    J Cell Biol. 1989 Jul;109(1):7-15 PMID: 2745558
  60. The coordinate release of cytochrome c during apoptosis is rapid, complete and kinetically invariant.
    Nat Cell Biol. 2000 Mar;2(3):156-62 PMID: 10707086
  61. Prostaglandins with antiproliferative activity induce the synthesis of a heat shock protein in human cells.
    Proc Natl Acad Sci U S A. 1989 Nov;86(21):8407-11 PMID: 2813398
  62. Induced thermotolerance to apoptosis in a human T lymphocyte cell line.
    J Cell Physiol. 1992 Jun;151(3):561-70 PMID: 1295903
  63. Heat shock protein hsp70 protects cells from thermal stress even after deletion of its ATP-binding domain.
    Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2036-40 PMID: 1549562
  64. BCL-2 family members and the mitochondria in apoptosis.
    Genes Dev. 1999 Aug 1;13(15):1899-911 PMID: 10444588
  65. Presence of a pre-apoptotic complex of pro-caspase-3, Hsp60 and Hsp10 in the mitochondrial fraction of jurkat cells.
    EMBO J. 1999 Apr 15;18(8):2040-8 PMID: 10205158
  66. Heat shock protein 72 modulates pathways of stress-induced apoptosis.
    J Biol Chem. 1998 Jul 3;273(27):17147-53 PMID: 9642282
  67. Inhibition of proliferation and induction of apoptosis by abrogation of heat-shock protein (HSP) 70 expression in tumor cells.
    Cancer Immunol Immunother. 1995 Feb;40(2):73-8 PMID: 7882385
  68. New adenovirus vectors for protein production and gene transfer.
    Cytotechnology. 1998 Nov;28(1-3):53-64 PMID: 19003407
  69. Hsp60 accelerates the maturation of pro-caspase-3 by upstream activator proteases during apoptosis.
    EMBO J. 1999 Apr 15;18(8):2049-56 PMID: 10205159
  70. Growth, metabolic, and antibody production kinetics of hybridoma cell culture: 1. Analysis of data from controlled batch reactors.
    Biotechnol Prog. 1991 Nov-Dec;7(6):471-80 PMID: 1367749
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.20.19.7146-7159.2000
Email
[email protected]
Published
2000-10-00
Pages
7146-59
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC86268
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.20.19.7146-7159.2000 PMC Full Text View journal details
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