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. 2003 Jun 1;372(Pt 2):359–369. doi: 10.1042/BJ20030201

c-Jun N-terminal kinase (JNK)-mediated modulation of brain mitochondria function: new target proteins for JNK signalling in mitochondrion-dependent apoptosis.

Hagen Schroeter 1, Clinton S Boyd 1, Ruhi Ahmed 1, Jeremy P E Spencer 1, Roger F Duncan 1, Catherine Rice-Evans 1, Enrique Cadenas 1
PMCID: PMC1223409  PMID: 12614194

Abstract

The molecular mechanisms underlying the initiation and control of the release of cytochrome c during mitochondrion-dependent apoptosis are thought to involve the phosphorylation of mitochondrial Bcl-2 and Bcl-x(L). Although the c-Jun N-terminal kinase (JNK) has been proposed to mediate the phosphorylation of Bcl-2/Bcl-x(L) the mechanisms linking the modification of these proteins and the release of cytochrome c remain to be elucidated. This study was aimed at establishing interdependency between JNK signalling and mitochondrial apoptosis. Using an experimental model consisting of isolated, bioenergetically competent rat brain mitochondria, these studies show that (i) JNK catalysed the phosphorylation of Bcl-2 and Bcl-x(L) as well as other mitochondrial proteins, as shown by two-dimensional isoelectric focusing/SDS/PAGE; (ii) JNK-induced cytochrome c release, in a process independent of the permeability transition of the inner mitochondrial membrane (imPT) and insensitive to cyclosporin A; (iii) JNK mediated a partial collapse of the mitochondrial inner-membrane potential (Deltapsim) in an imPT- and cyclosporin A-independent manner; and (iv) JNK was unable to induce imPT/swelling and did not act as a co-inducer, but as an inhibitor of Ca-induced imPT. The results are discussed with regard to the functional link between the Deltapsim and factors influencing the permeability transition of the inner and outer mitochondrial membranes. Taken together, JNK-dependent phosphorylation of mitochondrial proteins including, but not limited to, Bcl-2/Bcl-x(L) may represent a potential of the modulation of mitochondrial function during apoptosis.

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Selected References

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  1. Adams J. M., Cory S. The Bcl-2 protein family: arbiters of cell survival. Science. 1998 Aug 28;281(5381):1322–1326. doi: 10.1126/science.281.5381.1322. [DOI] [PubMed] [Google Scholar]
  2. Anderson M. F., Sims N. R. Improved recovery of highly enriched mitochondrial fractions from small brain tissue samples. Brain Res Brain Res Protoc. 2000 Feb;5(1):95–101. doi: 10.1016/s1385-299x(99)00060-4. [DOI] [PubMed] [Google Scholar]
  3. Antonsson B., Martinou J. C. The Bcl-2 protein family. Exp Cell Res. 2000 Apr 10;256(1):50–57. doi: 10.1006/excr.2000.4839. [DOI] [PubMed] [Google Scholar]
  4. Aronis Anna, Komarnitsky Rita, Shilo Shani, Tirosh Oren. Membrane depolarization of isolated rat liver mitochondria attenuates permeability transition pore opening and oxidant production. Antioxid Redox Signal. 2002 Aug;4(4):647–654. doi: 10.1089/15230860260220157. [DOI] [PubMed] [Google Scholar]
  5. Attalla H., Westberg J. A., Andersson L. C., Adlercreutz H., Mäkelä T. P. 2-Methoxyestradiol-induced phosphorylation of Bcl-2: uncoupling from JNK/SAPK activation. Biochem Biophys Res Commun. 1998 Jun 29;247(3):616–619. doi: 10.1006/bbrc.1998.8870. [DOI] [PubMed] [Google Scholar]
  6. Blagosklonny M. V., Chuman Y., Bergan R. C., Fojo T. Mitogen-activated protein kinase pathway is dispensable for microtubule-active drug-induced Raf-1/Bcl-2 phosphorylation and apoptosis in leukemia cells. Leukemia. 1999 Jul;13(7):1028–1036. doi: 10.1038/sj.leu.2401449. [DOI] [PubMed] [Google Scholar]
  7. Boveris A., Cadenas E. Mitochondrial production of hydrogen peroxide regulation by nitric oxide and the role of ubisemiquinone. IUBMB Life. 2000 Oct-Nov;50(4-5):245–250. doi: 10.1080/713803732. [DOI] [PubMed] [Google Scholar]
  8. Breitschopf K., Haendeler J., Malchow P., Zeiher A. M., Dimmeler S. Posttranslational modification of Bcl-2 facilitates its proteasome-dependent degradation: molecular characterization of the involved signaling pathway. Mol Cell Biol. 2000 Mar;20(5):1886–1896. doi: 10.1128/mcb.20.5.1886-1896.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Brenner C., Cadiou H., Vieira H. L., Zamzami N., Marzo I., Xie Z., Leber B., Andrews D., Duclohier H., Reed J. C. Bcl-2 and Bax regulate the channel activity of the mitochondrial adenine nucleotide translocator. Oncogene. 2000 Jan 20;19(3):329–336. doi: 10.1038/sj.onc.1203298. [DOI] [PubMed] [Google Scholar]
  10. Brown G. C., Borutaite V. Nitric oxide, mitochondria, and cell death. IUBMB Life. 2001 Sep-Nov;52(3-5):189–195. doi: 10.1080/15216540152845993. [DOI] [PubMed] [Google Scholar]
  11. Chang B. S., Minn A. J., Muchmore S. W., Fesik S. W., Thompson C. B. Identification of a novel regulatory domain in Bcl-X(L) and Bcl-2. EMBO J. 1997 Mar 3;16(5):968–977. doi: 10.1093/emboj/16.5.968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cheng E. H., Wei M. C., Weiler S., Flavell R. A., Mak T. W., Lindsten T., Korsmeyer S. J. BCL-2, BCL-X(L) sequester BH3 domain-only molecules preventing BAX- and BAK-mediated mitochondrial apoptosis. Mol Cell. 2001 Sep;8(3):705–711. doi: 10.1016/s1097-2765(01)00320-3. [DOI] [PubMed] [Google Scholar]
  13. Crompton M. Bax, Bid and the permeabilization of the mitochondrial outer membrane in apoptosis. Curr Opin Cell Biol. 2000 Aug;12(4):414–419. doi: 10.1016/s0955-0674(00)00110-1. [DOI] [PubMed] [Google Scholar]
  14. Crompton M. Mitochondrial intermembrane junctional complexes and their role in cell death. J Physiol. 2000 Nov 15;529(Pt 1):11–21. doi: 10.1111/j.1469-7793.2000.00011.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Davis R. J. Signal transduction by the JNK group of MAP kinases. Cell. 2000 Oct 13;103(2):239–252. doi: 10.1016/s0092-8674(00)00116-1. [DOI] [PubMed] [Google Scholar]
  16. Deng X., Xiao L., Lang W., Gao F., Ruvolo P., May W. S., Jr Novel role for JNK as a stress-activated Bcl2 kinase. J Biol Chem. 2001 Apr 25;276(26):23681–23688. doi: 10.1074/jbc.M100279200. [DOI] [PubMed] [Google Scholar]
  17. Diaz Brinton R., Chen S., Montoya M., Hsieh D., Minaya J., Kim J., Chu H. P. The women's health initiative estrogen replacement therapy is neurotrophic and neuroprotective. Neurobiol Aging. 2000 May-Jun;21(3):475–496. doi: 10.1016/s0197-4580(00)00109-3. [DOI] [PubMed] [Google Scholar]
  18. Duncan R., Hershey J. W. Evaluation of isoelectric focusing running conditions during two-dimensional isoelectric focusing/sodium dodecyl sulfate-polyacrylamide gel electrophoresis: variation of gel patterns with changing conditions and optimized isoelectric focusing conditions. Anal Biochem. 1984 Apr;138(1):144–155. doi: 10.1016/0003-2697(84)90783-8. [DOI] [PubMed] [Google Scholar]
  19. Dunkley P. R., Baker C. M., Robinson P. J. Depolarization-dependent protein phosphorylation in rat cortical synaptosomes: characterization of active protein kinases by phosphopeptide analysis of substrates. J Neurochem. 1986 Jun;46(6):1692–1703. doi: 10.1111/j.1471-4159.1986.tb08486.x. [DOI] [PubMed] [Google Scholar]
  20. Elfering Sarah Liv, Sarkela Theresa Marie, Giulivi Cecilia. Biochemistry of mitochondrial nitric-oxide synthase. J Biol Chem. 2002 Aug 1;277(41):38079–38086. doi: 10.1074/jbc.M205256200. [DOI] [PubMed] [Google Scholar]
  21. Fan M., Goodwin M., Vu T., Brantley-Finley C., Gaarde W. A., Chambers T. C. Vinblastine-induced phosphorylation of Bcl-2 and Bcl-XL is mediated by JNK and occurs in parallel with inactivation of the Raf-1/MEK/ERK cascade. J Biol Chem. 2000 Sep 29;275(39):29980–29985. doi: 10.1074/jbc.M003776200. [DOI] [PubMed] [Google Scholar]
  22. Gunter K. K., Gunter T. E. Transport of calcium by mitochondria. J Bioenerg Biomembr. 1994 Oct;26(5):471–485. doi: 10.1007/BF00762732. [DOI] [PubMed] [Google Scholar]
  23. Hengartner M. O. The biochemistry of apoptosis. Nature. 2000 Oct 12;407(6805):770–776. doi: 10.1038/35037710. [DOI] [PubMed] [Google Scholar]
  24. Ito T., Deng X., Carr B., May W. S. Bcl-2 phosphorylation required for anti-apoptosis function. J Biol Chem. 1997 May 2;272(18):11671–11673. doi: 10.1074/jbc.272.18.11671. [DOI] [PubMed] [Google Scholar]
  25. Jürgensmeier J. M., Xie Z., Deveraux Q., Ellerby L., Bredesen D., Reed J. C. Bax directly induces release of cytochrome c from isolated mitochondria. Proc Natl Acad Sci U S A. 1998 Apr 28;95(9):4997–5002. doi: 10.1073/pnas.95.9.4997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kahlert Stefan, Reiser Georg. Swelling of mitochondria in cultured rat hippocampal astrocytes is induced by high cytosolic Ca(2+) load, but not by mitochondrial depolarization. FEBS Lett. 2002 Oct 9;529(2-3):351–355. doi: 10.1016/s0014-5793(02)03394-x. [DOI] [PubMed] [Google Scholar]
  27. Kharbanda S., Saxena S., Yoshida K., Pandey P., Kaneki M., Wang Q., Cheng K., Chen Y. N., Campbell A., Sudha T. Translocation of SAPK/JNK to mitochondria and interaction with Bcl-x(L) in response to DNA damage. J Biol Chem. 2000 Jan 7;275(1):322–327. doi: 10.1074/jbc.275.1.322. [DOI] [PubMed] [Google Scholar]
  28. Lee Icksoo, Bender Elisabeth, Kadenbach Bernhard. Control of mitochondrial membrane potential and ROS formation by reversible phosphorylation of cytochrome c oxidase. Mol Cell Biochem. 2002 May-Jun;234-235(1-2):63–70. [PubMed] [Google Scholar]
  29. Lemeshko Victor V. Model of the outer membrane potential generation by the inner membrane of mitochondria. Biophys J. 2002 Feb;82(2):684–692. doi: 10.1016/S0006-3495(02)75431-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ling Y. H., Tornos C., Perez-Soler R. Phosphorylation of Bcl-2 is a marker of M phase events and not a determinant of apoptosis. J Biol Chem. 1998 Jul 24;273(30):18984–18991. doi: 10.1074/jbc.273.30.18984. [DOI] [PubMed] [Google Scholar]
  31. Martindale Jennifer L., Holbrook Nikki J. Cellular response to oxidative stress: signaling for suicide and survival. J Cell Physiol. 2002 Jul;192(1):1–15. doi: 10.1002/jcp.10119. [DOI] [PubMed] [Google Scholar]
  32. Maundrell K., Antonsson B., Magnenat E., Camps M., Muda M., Chabert C., Gillieron C., Boschert U., Vial-Knecht E., Martinou J. C. Bcl-2 undergoes phosphorylation by c-Jun N-terminal kinase/stress-activated protein kinases in the presence of the constitutively active GTP-binding protein Rac1. J Biol Chem. 1997 Oct 3;272(40):25238–25242. doi: 10.1074/jbc.272.40.25238. [DOI] [PubMed] [Google Scholar]
  33. Papa S., Scacco S., Sardanelli A. M., Petruzzella V., Vergari R., Signorile A., Technikova-Dobrova Z. Complex I and the cAMP cascade in human physiopathology. Biosci Rep. 2002 Feb;22(1):3–16. doi: 10.1023/a:1016004921277. [DOI] [PubMed] [Google Scholar]
  34. Park H. S., Park E., Kim M. S., Ahn K., Kim I. Y., Choi E. J. Selenite inhibits the c-Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK) through a thiol redox mechanism. J Biol Chem. 2000 Jan 28;275(4):2527–2531. doi: 10.1074/jbc.275.4.2527. [DOI] [PubMed] [Google Scholar]
  35. SWANSON M. A. Phosphatases of liver. I. Glucose-6-phosphatase. J Biol Chem. 1950 Jun;184(2):647–659. [PubMed] [Google Scholar]
  36. Scaduto R. C., Jr, Grotyohann L. W. Measurement of mitochondrial membrane potential using fluorescent rhodamine derivatives. Biophys J. 1999 Jan;76(1 Pt 1):469–477. doi: 10.1016/S0006-3495(99)77214-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Schroeter H., Williams R. J., Matin R., Iversen L., Rice-Evans C. A. Phenolic antioxidants attenuate neuronal cell death following uptake of oxidized low-density lipoprotein. Free Radic Biol Med. 2000 Dec 15;29(12):1222–1233. doi: 10.1016/s0891-5849(00)00415-9. [DOI] [PubMed] [Google Scholar]
  38. Shimizu S., Narita M., Tsujimoto Y. Bcl-2 family proteins regulate the release of apoptogenic cytochrome c by the mitochondrial channel VDAC. Nature. 1999 Jun 3;399(6735):483–487. doi: 10.1038/20959. [DOI] [PubMed] [Google Scholar]
  39. Sottocasa G. L., Kuylenstierna B., Ernster L., Bergstrand A. An electron-transport system associated with the outer membrane of liver mitochondria. A biochemical and morphological study. J Cell Biol. 1967 Feb;32(2):415–438. doi: 10.1083/jcb.32.2.415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Srivastava R. K., Mi Q. S., Hardwick J. M., Longo D. L. Deletion of the loop region of Bcl-2 completely blocks paclitaxel-induced apoptosis. Proc Natl Acad Sci U S A. 1999 Mar 30;96(7):3775–3780. doi: 10.1073/pnas.96.7.3775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Thompson C. B. Apoptosis in the pathogenesis and treatment of disease. Science. 1995 Mar 10;267(5203):1456–1462. doi: 10.1126/science.7878464. [DOI] [PubMed] [Google Scholar]
  42. Tournier C., Hess P., Yang D. D., Xu J., Turner T. K., Nimnual A., Bar-Sagi D., Jones S. N., Flavell R. A., Davis R. J. Requirement of JNK for stress-induced activation of the cytochrome c-mediated death pathway. Science. 2000 May 5;288(5467):870–874. doi: 10.1126/science.288.5467.870. [DOI] [PubMed] [Google Scholar]
  43. Tseng Chia-Jen, Wang Ying-Jan, Liang Yu-Chih, Jeng Jiiang-Huei, Lee Wen-Sen, Lin Jen-Kun, Chen Chien-Ho, Liu I-Chun, Ho Yuan-Soon. Microtubule damaging agents induce apoptosis in HL 60 cells and G2/M cell cycle arrest in HT 29 cells. Toxicology. 2002 Jun 14;175(1-3):123–142. doi: 10.1016/s0300-483x(02)00073-2. [DOI] [PubMed] [Google Scholar]
  44. Veis D. J., Sorenson C. M., Shutter J. R., Korsmeyer S. J. Bcl-2-deficient mice demonstrate fulminant lymphoid apoptosis, polycystic kidneys, and hypopigmented hair. Cell. 1993 Oct 22;75(2):229–240. doi: 10.1016/0092-8674(93)80065-m. [DOI] [PubMed] [Google Scholar]
  45. Wiltshire Carolyn, Matsushita Masato, Tsukada Satoshi, Gillespie David A. F., May Gerhard H. W. A new c-Jun N-terminal kinase (JNK)-interacting protein, Sab (SH3BP5), associates with mitochondria. Biochem J. 2002 Nov 1;367(Pt 3):577–585. doi: 10.1042/BJ20020553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Yamamoto K., Ichijo H., Korsmeyer S. J. BCL-2 is phosphorylated and inactivated by an ASK1/Jun N-terminal protein kinase pathway normally activated at G(2)/M. Mol Cell Biol. 1999 Dec;19(12):8469–8478. doi: 10.1128/mcb.19.12.8469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Yuan J., Yankner B. A. Apoptosis in the nervous system. Nature. 2000 Oct 12;407(6805):802–809. doi: 10.1038/35037739. [DOI] [PubMed] [Google Scholar]
  48. Zamzami N., Kroemer G. The mitochondrion in apoptosis: how Pandora's box opens. Nat Rev Mol Cell Biol. 2001 Jan;2(1):67–71. doi: 10.1038/35048073. [DOI] [PubMed] [Google Scholar]

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