Advertisement

Neurochemical Research

, Volume 32, Issue 9, pp 1573–1585 | Cite as

Nerve Growth Factor Potentiates p53 DNA Binding but Inhibits Nitric Oxide-Induced Apoptosis in Neuronal PC12 Cells

  • Christopher Brynczka
  • Bruce Alex Merrick
Original Paper

Abstract

NGF is recognized for its role in neuronal differentiation and maintenance. Differentiation of PC12 cells by NGF involves p53, a transcription factor that controls growth arrest and apoptosis. We investigated NGF influence over p53 activity during NO-induced apoptosis by sodium nitroprusside in differentiated and mitotic PC12 cells. NGF-differentiation produced increased p53 levels, nuclear localization and sequence-specific DNA binding. Apoptosis in mitotic cells also produced these events but the accompanying activation of caspases 1-10 and mitochondrial depolarization were inhibited during NGF differentiation and could be reversed in p53-silenced cells. Transcriptional regulation of PUMA and survivin expression were not inhibited by NGF, although NO-induced mitochondrial depolarization was dependent upon de novo gene transcription and only occurred in mitotic cells. We conclude that NGF mediates prosurvival signaling by increasing factors such as Bcl-2 and p21Waf1/Cip1 without altering p53 transcriptional activity to inhibit mitochondrial depolarization, caspase activation and apoptosis.

Keywords

NO PC12 Mitochondria Differentiation NGF p53 

Notes

Acknowledgements

This work was supported by the Intramural Research Program of the NIH, National Institute of Environmental Health Sciences. We thank Dr. Alexandra Heinloth and Dr. Sonnet Arlander for critical review of this manuscript.

References

  1. 1.
    Kuida K, Zheng TS, Na S et al (1996) Decreased apoptosis in the brain and premature lethality in CPP32-deficient mice. Nature 384:368–372PubMedCrossRefGoogle Scholar
  2. 2.
    Mattson MP (2000) Apoptosis in neurodegenerative disorders. Nat Rev Mol Cell Biol 1:120–129PubMedCrossRefGoogle Scholar
  3. 3.
    Vousden KH, Lu X (2002) Live or let die: the cell’s response to p53. Nat Rev Cancer 2:594–604PubMedCrossRefGoogle Scholar
  4. 4.
    Yee KS, Vousden KH (2005) Complicating the complexity of p53. Carcinogenesis 26:1317–1322PubMedCrossRefGoogle Scholar
  5. 5.
    Chipuk JE, Kuwana T, Bouchier-Hayes L et al (2004) Direct activation of Bax by p53 mediates mitochondrial membrane permeabilization and apoptosis. Science 303:1010–1014PubMedCrossRefGoogle Scholar
  6. 6.
    Chipuk JE, Bouchier-Hayes L, Green DR (2006) Mitochondrial outer membrane permeabilization during apoptosis: the innocent bystander scenario. Cell Death Differ 13:1396–1402PubMedCrossRefGoogle Scholar
  7. 7.
    Zhang J, Yan W, Chen X (2006) p53 is required for nerve growth factor-mediated differentiation of PC12 cells via regulation of TrkA levels. Cell Death Differ 13:2118–2128PubMedCrossRefGoogle Scholar
  8. 8.
    Di Giovanni S, Knights CD, Rao M et al (2006) The tumor suppressor protein p53 is required for neurite outgrowth and axon regeneration. Embo J 25:4084–4096PubMedCrossRefGoogle Scholar
  9. 9.
    Hughes AL, Gollapudi L, Sladek TL et al (2000) Mediation of nerve growth factor-driven cell cycle arrest in PC12 cells by p53. Simultaneous differentiation and proliferation subsequent to p53 functional inactivation. J Biol Chem 275:37829–37837PubMedCrossRefGoogle Scholar
  10. 10.
    Klein R, Jing SQ, Nanduri V et al (1991) The trk proto-oncogene encodes a receptor for nerve growth factor. Cell 65:189–197PubMedCrossRefGoogle Scholar
  11. 11.
    Greene LA, Tischler AS (1976) Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc Natl Acad Sci USA 73:2424–2428PubMedCrossRefGoogle Scholar
  12. 12.
    Sofroniew MV, Howe CL, Mobley WC (2001) Nerve growth factor signaling, neuroprotection, and neural repair. Annu Rev Neurosci 24:1217–1281PubMedCrossRefGoogle Scholar
  13. 13.
    Andjelkovic M, Suidan HS, Meier R et al (1998) Nerve growth factor promotes activation of the alpha, beta and gamma isoforms of protein kinase B in PC12 pheochromocytoma cells. Eur J Biochem 251:195–200PubMedCrossRefGoogle Scholar
  14. 14.
    Crowder RJ, Freeman RS (1998) Phosphatidylinositol 3-kinase and Akt protein kinase are necessary and sufficient for the survival of nerve growth factor-dependent sympathetic neurons. J Neurosci 18:2933–2943PubMedGoogle Scholar
  15. 15.
    Xue L, Murray JH, Tolkovsky AM (2000) The Ras/phosphatidylinositol 3-kinase and Ras/ERK pathways function as independent survival modules each of which inhibits a distinct apoptotic signaling pathway in sympathetic neurons. J Biol Chem 275:8817–8824PubMedCrossRefGoogle Scholar
  16. 16.
    Liu H, Nowak R, Chao W et al (2003) Nerve growth factor induces anti-apoptotic heme oxygenase-1 in rat pheochromocytoma PC12 cells. J Neurochem 86:1553–1563PubMedCrossRefGoogle Scholar
  17. 17.
    Salinas M, Diaz R, Abraham NG et al (2003) Nerve growth factor protects against 6-hydroxydopamine-induced oxidative stress by increasing expression of heme oxygenase-1 in a phosphatidylinositol 3-kinase-dependent manner. J Biol Chem 278:13898–13904PubMedCrossRefGoogle Scholar
  18. 18.
    Ekshyyan O, Aw TY (2005) Decreased susceptibility of differentiated PC12 cells to oxidative challenge: relationship to cellular redox and expression of apoptotic protease activator factor-1. Cell Death Differ 12:1066–1077PubMedCrossRefGoogle Scholar
  19. 19.
    Vyas S, Juin P, Hancock D et al (2004) Differentiation-dependent sensitivity to apoptogenic factors in PC12 cells. J Biol Chem 279:30983–30993PubMedCrossRefGoogle Scholar
  20. 20.
    Wada K, Okada N, Yamamura T et al (1996) Nerve growth factor induces resistance of PC12 cells to nitric oxide cytotoxicity. Neurochem Int 29:461–467PubMedCrossRefGoogle Scholar
  21. 21.
    Liaudet L, Soriano FG, Szabo C (2000) Biology of nitric oxide signaling. Crit Care Med 28:N37–N52PubMedCrossRefGoogle Scholar
  22. 22.
    Phung YT, Bekker JM, Hallmark OG et al (1999) Both neuronal NO synthase and nitric oxide are required for PC12 cell differentiation: a cGMP independent pathway. Brain Res Mol Brain Res 64:165–178PubMedCrossRefGoogle Scholar
  23. 23.
    Bredt DS, Hwang PM, Snyder SH (1990) Localization of nitric oxide synthase indicating a neural role for nitric oxide. Nature 347:768–770PubMedCrossRefGoogle Scholar
  24. 24.
    Blaise GA, Gauvin D, Gangal M et al (2005) Nitric oxide, cell signaling and cell death. Toxicology 208:177–192PubMedCrossRefGoogle Scholar
  25. 25.
    Farinelli SE, Park DS, Greene LA (1996) Nitric oxide delays the death of trophic factor-deprived PC12 cells and sympathetic neurons by a cGMP-mediated mechanism. J Neurosci 16:2325–2334PubMedGoogle Scholar
  26. 26.
    Heinloth A, Brune B, Fischer B et al (2002) Nitric oxide prevents oxidised LDL-induced p53 accumulation, cytochrome c translocation, and apoptosis in macrophages via guanylate cyclase stimulation. Atherosclerosis 162:93–101PubMedCrossRefGoogle Scholar
  27. 27.
    Fraser M, Chan SL, Chan SS et al (2006) Regulation of p53 and suppression of apoptosis by the soluble guanylyl cyclase/cGMP pathway in human ovarian cancer cells. Oncogene 25:2203–2212PubMedCrossRefGoogle Scholar
  28. 28.
    Torok NJ, Higuchi H, Bronk S et al (2002) Nitric oxide inhibits apoptosis downstream of cytochrome C release by nitrosylating caspase 9. Cancer Res 62:1648–1653PubMedGoogle Scholar
  29. 29.
    Schneiderhan N, Budde A, Zhang Y et al (2003) Nitric oxide induces phosphorylation of p53 and impairs nuclear export. Oncogene 22:2857–2868PubMedCrossRefGoogle Scholar
  30. 30.
    Yung HW, Bal-Price AK, Brown GC et al (2004) Nitric oxide-induced cell death of cerebrocortical murine astrocytes is mediated through p53- and Bax-dependent pathways. J Neurochem 89:812–821PubMedCrossRefGoogle Scholar
  31. 31.
    Li CQ, Trudel LJ, Wogan GN (2002) Nitric oxide-induced genotoxicity, mitochondrial damage, and apoptosis in human lymphoblastoid cells expressing wild-type and mutant p53. Proc Natl Acad Sci USA 99:10364–10369PubMedCrossRefGoogle Scholar
  32. 32.
    Hirsch EC, Hunot S, Damier P et al (1998) Glial cells and inflammation in Parkinson’s disease: a role in neurodegeneration? Ann Neurol 44:S115–S120PubMedGoogle Scholar
  33. 33.
    Liu B, Hong JS (2003) Role of microglia in inflammation-mediated neurodegenerative diseases: mechanisms and strategies for therapeutic intervention. J Pharmacol Exp Ther 304:1–7PubMedCrossRefGoogle Scholar
  34. 34.
    Gibbons HM, Dragunow M (2006) Microglia induce neural cell death via a proximity-dependent mechanism involving nitric oxide. Brain Res 1084:1–15PubMedCrossRefGoogle Scholar
  35. 35.
    Arimoto T, Bing G (2003) Up-regulation of inducible nitric oxide synthase in the substantia nigra by lipopolysaccharide causes microglial activation and neurodegeneration. Neurobiol Dis 12:35–45PubMedCrossRefGoogle Scholar
  36. 36.
    Liu B, Gao HM, Wang JY et al (2002) Role of nitric oxide in inflammation-mediated neurodegeneration. Ann NY Acad Sci 962:318–331PubMedCrossRefGoogle Scholar
  37. 37.
    Bal-Price A, Brown GC (2000) Nitric-oxide-induced necrosis and apoptosis in PC12 cells mediated by mitochondria. J Neurochem 75:1455–1464PubMedCrossRefGoogle Scholar
  38. 38.
    McNeill-Blue C, Wetmore BA, Sanchez JF et al (2006) Apoptosis mediated by p53 in rat neural AF5 cells following treatment with hydrogen peroxide and staurosporine. Brain Res 1112:1–15PubMedCrossRefGoogle Scholar
  39. 39.
    Labhart P, Karmakar S, Salicru EM et al (2005) Identification of target genes in breast cancer cells directly regulated by the SRC-3/AIB1 coactivator. Proc Natl Acad Sci USA 102:1339–1344PubMedCrossRefGoogle Scholar
  40. 40.
    Soutoglou E, Talianidis I (2002) Coordination of PIC assembly and chromatin remodeling during differentiation-induced gene activation. Science 295:1901–1904PubMedCrossRefGoogle Scholar
  41. 41.
    Shieh SY, Ikeda M, Taya Y et al (1997) DNA damage-induced phosphorylation of p53 alleviates inhibition by MDM2. Cell 91:325–334PubMedCrossRefGoogle Scholar
  42. 42.
    Vicente S, Perez-Rodriguez R, Olivan AM et al (2006) Nitric oxide and peroxynitrite induce cellular death in bovine chromaffin cells: evidence for a mixed necrotic and apoptotic mechanism with caspases activation. J Neurosci Res 84:78–96PubMedCrossRefGoogle Scholar
  43. 43.
    Yu J, Zhang L (2005) The transcriptional targets of p53 in apoptosis control. Biochem Biophys Res Commun 331:851–858PubMedCrossRefGoogle Scholar
  44. 44.
    Hoffman WH, Biade S, Zilfou JT et al (2002) Transcriptional repression of the anti-apoptotic survivin gene by wild type p53. J Biol Chem 277:3247–3257PubMedCrossRefGoogle Scholar
  45. 45.
    Mirza A, McGuirk M, Hockenberry TN et al (2002) Human survivin is negatively regulated by wild-type p53 and participates in p53-dependent apoptotic pathway. Oncogene 21:2613–2622PubMedCrossRefGoogle Scholar
  46. 46.
    Nakaso K, Yoshimoto Y, Yano H et al (2004) p53-mediated mitochondrial dysfunction by proteasome inhibition in dopaminergic SH-SY5Y cells. Neurosci Lett 354:213–216PubMedCrossRefGoogle Scholar
  47. 47.
    Wang C, Trudel LJ, Wogan GN et al (2003) Thresholds of nitric oxide-mediated toxicity in human lymphoblastoid cells. Chem Res Toxicol 16:1004–1013PubMedCrossRefGoogle Scholar
  48. 48.
    Pallis M, Grundy M, Turzanski J et al (2001) Mitochondrial membrane sensitivity to depolarization in acute myeloblastic leukemia is associated with spontaneous in vitro apoptosis, wild-type TP53, and vicinal thiol/disulfide status. Blood 98:405–413PubMedCrossRefGoogle Scholar
  49. 49.
    Smaili SS, Hsu YT, Sanders KM et al (2001) Bax translocation to mitochondria subsequent to a rapid loss of mitochondrial membrane potential. Cell Death Differ 8:909–920PubMedCrossRefGoogle Scholar
  50. 50.
    Li MH, Jang JH, Surh YJ (2005) Nitric oxide induces apoptosis via AP-1-driven upregulation of COX-2 in rat pheochromocytoma cells. Free Radic Biol Med 39:890–899PubMedCrossRefGoogle Scholar
  51. 51.
    Katoh S, Mitsui Y, Kitani K et al (1996) Nerve growth factor rescues PC12 cells from apoptosis by increasing amount of bcl-2. Biochem Biophys Res Commun 229:653–657PubMedCrossRefGoogle Scholar
  52. 52.
    Dispersyn G, Nuydens R, Connors R et al (1999) Bcl-2 protects against FCCP-induced apoptosis and mitochondrial membrane potential depolarization in PC12 cells. Biochim Biophys Acta 1428:357–371PubMedGoogle Scholar
  53. 53.
    Armstrong JS, Steinauer KK, French J et al (2001) Bcl-2 inhibits apoptosis induced by mitochondrial uncoupling but does not prevent mitochondrial transmembrane depolarization. Exp Cell Res 262:170–179PubMedCrossRefGoogle Scholar
  54. 54.
    el-Deiry WS, Harper JW, O’Connor PM et al (1994) WAF1/CIP1 is induced in p53-mediated G1 arrest and apoptosis. Cancer Res 54:1169–1174PubMedGoogle Scholar
  55. 55.
    Sohn D, Essmann F, Schulze-Osthoff K et al (2006) p21 blocks irradiation-induced apoptosis downstream of mitochondria by inhibition of cyclin-dependent kinase-mediated caspase-9 activation. Cancer Res 66:11254–11262PubMedCrossRefGoogle Scholar
  56. 56.
    Javelaud D, Besancon F (2002) Inactivation of p21WAF1 sensitizes cells to apoptosis via an increase of both p14ARF and p53 levels and an alteration of the Bax/Bcl-2 ratio. J Biol Chem 277:37949–37954PubMedCrossRefGoogle Scholar
  57. 57.
    Gollapudi L, Neet KE (1997) Different mechanisms for inhibition of cell proliferation via cell cycle proteins in PC12 cells by nerve growth factor and staurosporine. J Neurosci Res 49:461–474PubMedCrossRefGoogle Scholar
  58. 58.
    Schlossmann J, Feil R, Hofmann F (2003) Signaling through NO and cGMP-dependent protein kinases. Ann Med 35:21–27PubMedCrossRefGoogle Scholar
  59. 59.
    Sun J, Steenbergen C, Murphy E (2006) S-nitrosylation: NO-related redox signaling to protect against oxidative stress. Antioxid Redox Signal 8:1693–1705PubMedCrossRefGoogle Scholar
  60. 60.
    Ischiropoulos H (2003) Biological selectivity and functional aspects of protein tyrosine nitration. Biochem Biophys Res Commun 305:776–783PubMedCrossRefGoogle Scholar
  61. 61.
    Li J, Billiar TR, Talanian RV et al (1997) Nitric oxide reversibly inhibits seven members of the caspase family via S-nitrosylation. Biochem Biophys Res Commun 240:419–424PubMedCrossRefGoogle Scholar
  62. 62.
    Li MH, Cha YN, Surh YJ (2006) Carbon monoxide protects PC12 cells from peroxynitrite-induced apoptotic death by preventing the depolarization of mitochondrial transmembrane potential. Biochem Biophys Res Commun 342:984–990PubMedCrossRefGoogle Scholar
  63. 63.
    Reiter TA, Pang B, Dedon P et al (2006) Resistance to nitric oxide-induced necrosis in heme oxygenase-1 overexpressing pulmonary epithelial cells associated with decreased lipid peroxidation. J Biol Chem 281:36603–36612PubMedCrossRefGoogle Scholar
  64. 64.
    Kim Y, Seger R, Suresh Babu CV et al (2004) A positive role of the PI3-K/Akt signaling pathway in PC12 cell differentiation. Mol Cells 18:353–359PubMedGoogle Scholar
  65. 65.
    Jackson TR, Blader IJ, Hammonds-Odie LP et al (1996) Initiation and maintenance of NGF-stimulated neurite outgrowth requires activation of a phosphoinositide 3-kinase. J Cell Sci 109( Pt 2):289–300PubMedGoogle Scholar
  66. 66.
    Ulrich E, Duwel A, Kauffmann-Zeh A et al (1998) Specific TrkA survival signals interfere with different apoptotic pathways. Oncogene 16:825–832PubMedCrossRefGoogle Scholar
  67. 67.
    Ahn JY, Liu X, Liu Z et al (2006) Nuclear Akt associates with PKC-phosphorylated Ebp1, preventing DNA fragmentation by inhibition of caspase-activated DNase. Embo J 25:2083–2095PubMedCrossRefGoogle Scholar
  68. 68.
    Pugazhenthi S, Nesterova A, Sable C et al (2000) Akt/protein kinase B up-regulates Bcl-2 expression through cAMP-response element-binding protein. J Biol Chem 275:10761–10766PubMedCrossRefGoogle Scholar
  69. 69.
    Hock C, Heese K, Hulette C et al (2000) Region-specific neurotrophin imbalances in Alzheimer disease: decreased levels of brain-derived neurotrophic factor and increased levels of nerve growth factor in hippocampus and cortical areas. Arch Neurol 57:846–851PubMedCrossRefGoogle Scholar
  70. 70.
    Mogi M, Togari A, Kondo T et al (1999) Brain-derived growth factor and nerve growth factor concentrations are decreased in the substantia nigra in Parkinson’s disease. Neurosci Lett 270:45–48PubMedCrossRefGoogle Scholar
  71. 71.
    Llado J, Haenggeli C, Maragakis NJ et al (2004) Neural stem cells protect against glutamate-induced excitotoxicity and promote survival of injured motor neurons through the secretion of neurotrophic factors. Mol Cell Neurosci 27:322–331PubMedGoogle Scholar
  72. 72.
    Lu P, Jones LL, Snyder EY et al (2003) Neural stem cells constitutively secrete neurotrophic factors and promote extensive host axonal growth after spinal cord injury. Exp Neurol 181:115–129PubMedCrossRefGoogle Scholar
  73. 73.
    Longo FM, Massa SM (2005) Neurotrophin receptor-based strategies for Alzheimer’s disease. Curr Alzheimer Res 2:167–169PubMedCrossRefGoogle Scholar
  74. 74.
    Schweigreiter R (2006) The dual nature of neurotrophins. Bioessays 28:583–594PubMedCrossRefGoogle Scholar
  75. 75.
    Cernak I, Stoica B, Byrnes KR et al (2005) Role of the cell cycle in the pathobiology of central nervous system trauma. Cell Cycle 4:1286–1293PubMedGoogle Scholar
  76. 76.
    Fishel ML, Vasko MR, Kelley MR (2007) DNA repair in neurons: So if they don’t divide what’s to repair? Mutat Res 614:24–36PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  1. 1.National Center for ToxicogenomicsNational Institute of Environmental Health Sciences, National Institutes of HealthResearch Triangle ParkUSA
  2. 2.Department of Environmental and Molecular ToxicologyNorth Carolina State UniversityRaleighUSA

Personalised recommendations