Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 2003 Aug 1;373(Pt 3):739–746. doi: 10.1042/BJ20021918

The transcription factor early growth response factor-1 (EGR-1) promotes apoptosis of neuroblastoma cells.

Miguel Pignatelli 1, Rosario Luna-Medina 1, Arturo Pérez-Rendón 1, Angel Santos 1, Ana Perez-Castillo 1
PMCID: PMC1223559  PMID: 12755686

Abstract

Early growth response factor-1 (EGR-1) is an immediate early gene, which is rapidly activated in quiescent cells by mitogens or in postmitotic neurons after depolarization. EGR-1 has been involved in diverse biological functions such as cell growth, differentiation and apoptosis. Here we report that enforced expression of the EGR-1 gene induces apoptosis, as determined by flow cytometry and terminal deoxynucleotidyl transferase-mediated dUTP-fluorescein nick-end labelling (TUNEL) analysis, in murine Neuro2A cells. In accordance with this role of EGR-1 in cell death, antisense oligonucleotides increase cell viability in cells cultured in the absence of serum. This apoptotic activity of the EGR-1 appears to be mediated by p73, a member of the p53 family of proteins, since an increase in the amount of p73 is observed in clones stably expressing the EGR-1 protein. We also observed an increase in the transcriptional activity of the mdm2 promoter in cells overexpressing EGR-1, which is paralleled by a marked decrease in the levels of p53 protein, therefore excluding a role of this protein in mediating EGR-1-induced apoptosis. Our results suggest that EGR-1 is an important factor involved in neuronal apoptosis.

Full Text

The Full Text of this article is available as a PDF (321.1 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Agami R., Blandino G., Oren M., Shaul Y. Interaction of c-Abl and p73alpha and their collaboration to induce apoptosis. Nature. 1999 Jun 24;399(6738):809–813. doi: 10.1038/21697. [DOI] [PubMed] [Google Scholar]
  2. Ahmed M. M., Venkatasubbarao K., Fruitwala S. M., Muthukkumar S., Wood D. P., Jr, Sells S. F., Mohiuddin M., Rangnekar V. M. EGR-1 induction is required for maximal radiosensitivity in A375-C6 melanoma cells. J Biol Chem. 1996 Nov 15;271(46):29231–29237. doi: 10.1074/jbc.271.46.29231. [DOI] [PubMed] [Google Scholar]
  3. Ahmed N. N., Grimes H. L., Bellacosa A., Chan T. O., Tsichlis P. N. Transduction of interleukin-2 antiapoptotic and proliferative signals via Akt protein kinase. Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):3627–3632. doi: 10.1073/pnas.94.8.3627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Barres B. A., Raff M. C., Gaese F., Bartke I., Dechant G., Barde Y. A. A crucial role for neurotrophin-3 in oligodendrocyte development. Nature. 1994 Jan 27;367(6461):371–375. doi: 10.1038/367371a0. [DOI] [PubMed] [Google Scholar]
  5. Bartholomä Pia, Erlandsson Nina, Kaufmann Katrin, Rössler Oliver G., Baumann Bernd, Wirth Thomas, Giehl Klaus M., Thiel Gerald. Neuronal cell death induced by antidepressants: lack of correlation with Egr-1, NF-kappa B and extracellular signal-regulated protein kinase activation. Biochem Pharmacol. 2002 Apr 15;63(8):1507–1516. doi: 10.1016/s0006-2952(02)00882-1. [DOI] [PubMed] [Google Scholar]
  6. Beckmann A. M., Wilce P. A. Egr transcription factors in the nervous system. Neurochem Int. 1997 Oct;31(4):477–526. doi: 10.1016/s0197-0186(96)00136-2. [DOI] [PubMed] [Google Scholar]
  7. Brown D. R., Deb S., Muñoz R. M., Subler M. A., Deb S. P. The tumor suppressor p53 and the oncoprotein simian virus 40 T antigen bind to overlapping domains on the MDM2 protein. Mol Cell Biol. 1993 Nov;13(11):6849–6857. doi: 10.1128/mcb.13.11.6849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brummelkamp Thijn R., Bernards René, Agami Reuven. A system for stable expression of short interfering RNAs in mammalian cells. Science. 2002 Mar 21;296(5567):550–553. doi: 10.1126/science.1068999. [DOI] [PubMed] [Google Scholar]
  9. Catania M. V., Copani A., Calogero A., Ragonese G. I., Condorelli D. F., Nicoletti F. An enhanced expression of the immediate early gene, Egr-1, is associated with neuronal apoptosis in culture. Neuroscience. 1999;91(4):1529–1538. doi: 10.1016/s0306-4522(98)00544-2. [DOI] [PubMed] [Google Scholar]
  10. Catania M. V., Copani A., Calogero A., Ragonese G. I., Condorelli D. F., Nicoletti F. An enhanced expression of the immediate early gene, Egr-1, is associated with neuronal apoptosis in culture. Neuroscience. 1999;91(4):1529–1538. doi: 10.1016/s0306-4522(98)00544-2. [DOI] [PubMed] [Google Scholar]
  11. Chen J., Wu X., Lin J., Levine A. J. mdm-2 inhibits the G1 arrest and apoptosis functions of the p53 tumor suppressor protein. Mol Cell Biol. 1996 May;16(5):2445–2452. doi: 10.1128/mcb.16.5.2445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cibelli Giuseppe, Policastro Vincenza, Rössler Oliver G., Thiel Gerald. Nitric oxide-induced programmed cell death in human neuroblastoma cells is accompanied by the synthesis of Egr-1, a zinc finger transcription factor. J Neurosci Res. 2002 Feb 15;67(4):450–460. doi: 10.1002/jnr.10141. [DOI] [PubMed] [Google Scholar]
  13. Connor B., Dragunow M. The role of neuronal growth factors in neurodegenerative disorders of the human brain. Brain Res Brain Res Rev. 1998 Jun;27(1):1–39. doi: 10.1016/s0165-0173(98)00004-6. [DOI] [PubMed] [Google Scholar]
  14. Crosby S. D., Puetz J. J., Simburger K. S., Fahrner T. J., Milbrandt J. The early response gene NGFI-C encodes a zinc finger transcriptional activator and is a member of the GCGGGGGCG (GSG) element-binding protein family. Mol Cell Biol. 1991 Aug;11(8):3835–3841. doi: 10.1128/mcb.11.8.3835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Davis P. K., Dowdy S. F. p73. Int J Biochem Cell Biol. 2001 Oct;33(10):935–939. doi: 10.1016/s1357-2725(01)00073-5. [DOI] [PubMed] [Google Scholar]
  16. De Laurenzi V., Raschellá G., Barcaroli D., Annicchiarico-Petruzzelli M., Ranalli M., Catani M. V., Tanno B., Costanzo A., Levrero M., Melino G. Induction of neuronal differentiation by p73 in a neuroblastoma cell line. J Biol Chem. 2000 May 19;275(20):15226–15231. doi: 10.1074/jbc.275.20.15226. [DOI] [PubMed] [Google Scholar]
  17. Dragunow M. A role for immediate-early transcription factors in learning and memory. Behav Genet. 1996 May;26(3):293–299. doi: 10.1007/BF02359385. [DOI] [PubMed] [Google Scholar]
  18. Dussault J. H., Ruel J. Thyroid hormones and brain development. Annu Rev Physiol. 1987;49:321–334. doi: 10.1146/annurev.ph.49.030187.001541. [DOI] [PubMed] [Google Scholar]
  19. Estus S., Zaks W. J., Freeman R. S., Gruda M., Bravo R., Johnson E. M., Jr Altered gene expression in neurons during programmed cell death: identification of c-jun as necessary for neuronal apoptosis. J Cell Biol. 1994 Dec;127(6 Pt 1):1717–1727. doi: 10.1083/jcb.127.6.1717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ferreira A., Kosik K. S. Accelerated neuronal differentiation induced by p53 suppression. J Cell Sci. 1996 Jun;109(Pt 6):1509–1516. doi: 10.1242/jcs.109.6.1509. [DOI] [PubMed] [Google Scholar]
  21. Garcia-Valenzuela E., Gorczyca W., Darzynkiewicz Z., Sharma S. C. Apoptosis in adult retinal ganglion cells after axotomy. J Neurobiol. 1994 Apr;25(4):431–438. doi: 10.1002/neu.480250408. [DOI] [PubMed] [Google Scholar]
  22. Haines D. S., Landers J. E., Engle L. J., George D. L. Physical and functional interaction between wild-type p53 and mdm2 proteins. Mol Cell Biol. 1994 Feb;14(2):1171–1178. doi: 10.1128/mcb.14.2.1171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ham J., Babij C., Whitfield J., Pfarr C. M., Lallemand D., Yaniv M., Rubin L. L. A c-Jun dominant negative mutant protects sympathetic neurons against programmed cell death. Neuron. 1995 May;14(5):927–939. doi: 10.1016/0896-6273(95)90331-3. [DOI] [PubMed] [Google Scholar]
  24. Haupt Y., Maya R., Kazaz A., Oren M. Mdm2 promotes the rapid degradation of p53. Nature. 1997 May 15;387(6630):296–299. doi: 10.1038/387296a0. [DOI] [PubMed] [Google Scholar]
  25. Jensen P. J., Millan N., Mack K. J. Cortical NMDAR-1 gene expression is rapidly upregulated after seizure. Brain Res Mol Brain Res. 1997 Feb;44(1):157–162. doi: 10.1016/s0169-328x(96)00262-8. [DOI] [PubMed] [Google Scholar]
  26. Johnson E. M., Jr, Chang J. Y., Koike T., Martin D. P. Why do neurons die when deprived of trophic factor? Neurobiol Aging. 1989 Sep-Oct;10(5):549–553. doi: 10.1016/0197-4580(89)90127-9. [DOI] [PubMed] [Google Scholar]
  27. Juven T., Barak Y., Zauberman A., George D. L., Oren M. Wild type p53 can mediate sequence-specific transactivation of an internal promoter within the mdm2 gene. Oncogene. 1993 Dec;8(12):3411–3416. [PubMed] [Google Scholar]
  28. Kaetzel D. M., Jr, Maul R. S., Liu B., Bonthron D., Fenstermaker R. A., Coyne D. W. Platelet-derived growth factor A-chain gene transcription is mediated by positive and negative regulatory regions in the promoter. Biochem J. 1994 Jul 15;301(Pt 2):321–327. doi: 10.1042/bj3010321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kern S. E., Pietenpol J. A., Thiagalingam S., Seymour A., Kinzler K. W., Vogelstein B. Oncogenic forms of p53 inhibit p53-regulated gene expression. Science. 1992 May 8;256(5058):827–830. doi: 10.1126/science.1589764. [DOI] [PubMed] [Google Scholar]
  30. Krishnaraju K., Nguyen H. Q., Liebermann D. A., Hoffman B. The zinc finger transcription factor Egr-1 potentiates macrophage differentiation of hematopoietic cells. Mol Cell Biol. 1995 Oct;15(10):5499–5507. doi: 10.1128/mcb.15.10.5499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Kubbutat M. H., Jones S. N., Vousden K. H. Regulation of p53 stability by Mdm2. Nature. 1997 May 15;387(6630):299–303. doi: 10.1038/387299a0. [DOI] [PubMed] [Google Scholar]
  32. Lemaire P., Vesque C., Schmitt J., Stunnenberg H., Frank R., Charnay P. The serum-inducible mouse gene Krox-24 encodes a sequence-specific transcriptional activator. Mol Cell Biol. 1990 Jul;10(7):3456–3467. doi: 10.1128/mcb.10.7.3456. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Levkovitz Y., Baraban J. M. A dominant negative inhibitor of the Egr family of transcription regulatory factors suppresses cerebellar granule cell apoptosis by blocking c-Jun activation. J Neurosci. 2001 Aug 15;21(16):5893–5901. doi: 10.1523/JNEUROSCI.21-16-05893.2001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Levrero M., De Laurenzi V., Costanzo A., Gong J., Wang J. Y., Melino G. The p53/p63/p73 family of transcription factors: overlapping and distinct functions. J Cell Sci. 2000 May;113(Pt 10):1661–1670. doi: 10.1242/jcs.113.10.1661. [DOI] [PubMed] [Google Scholar]
  35. Li Y., Camp S., Rachinsky T. L., Bongiorno C., Taylor P. Promoter elements and transcriptional control of the mouse acetylcholinesterase gene. J Biol Chem. 1993 Feb 15;268(5):3563–3572. [PubMed] [Google Scholar]
  36. Liu C., Rangnekar V. M., Adamson E., Mercola D. Suppression of growth and transformation and induction of apoptosis by EGR-1. Cancer Gene Ther. 1998 Jan-Feb;5(1):3–28. [PubMed] [Google Scholar]
  37. Louis J. C., Magal E., Takayama S., Varon S. CNTF protection of oligodendrocytes against natural and tumor necrosis factor-induced death. Science. 1993 Jan 29;259(5095):689–692. doi: 10.1126/science.8430320. [DOI] [PubMed] [Google Scholar]
  38. McKinnon R. D., Matsui T., Dubois-Dalcq M., Aaronson S. A. FGF modulates the PDGF-driven pathway of oligodendrocyte development. Neuron. 1990 Nov;5(5):603–614. doi: 10.1016/0896-6273(90)90215-2. [DOI] [PubMed] [Google Scholar]
  39. Mellström B., Pipaón C., Naranjo J. R., Perez-Castillo A., Santos A. Differential effect of thyroid hormone on NGFI-A gene expression in developing rat brain. Endocrinology. 1994 Aug;135(2):583–588. doi: 10.1210/endo.135.2.8033806. [DOI] [PubMed] [Google Scholar]
  40. Menéndez-Hurtado A., Santos A., Pérez-Castillo A. Characterization of the promoter region of the rat CCAAT/enhancer-binding protein alpha gene and regulation by thyroid hormone in rat immortalized brown adipocytes. Endocrinology. 2000 Nov;141(11):4164–4170. doi: 10.1210/endo.141.11.7756. [DOI] [PubMed] [Google Scholar]
  41. Milbrandt J. A nerve growth factor-induced gene encodes a possible transcriptional regulatory factor. Science. 1987 Nov 6;238(4828):797–799. doi: 10.1126/science.3672127. [DOI] [PubMed] [Google Scholar]
  42. Molnar G., Crozat A., Pardee A. B. The immediate-early gene Egr-1 regulates the activity of the thymidine kinase promoter at the G0-to-G1 transition of the cell cycle. Mol Cell Biol. 1994 Aug;14(8):5242–5248. doi: 10.1128/mcb.14.8.5242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Muthukkumar S., Nair P., Sells S. F., Maddiwar N. G., Jacob R. J., Rangnekar V. M. Role of EGR-1 in thapsigargin-inducible apoptosis in the melanoma cell line A375-C6. Mol Cell Biol. 1995 Nov;15(11):6262–6272. doi: 10.1128/mcb.15.11.6262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Nair P., Muthukkumar S., Sells S. F., Han S. S., Sukhatme V. P., Rangnekar V. M. Early growth response-1-dependent apoptosis is mediated by p53. J Biol Chem. 1997 Aug 8;272(32):20131–20138. doi: 10.1074/jbc.272.32.20131. [DOI] [PubMed] [Google Scholar]
  45. Nguyen H. Q., Hoffman-Liebermann B., Liebermann D. A. The zinc finger transcription factor Egr-1 is essential for and restricts differentiation along the macrophage lineage. Cell. 1993 Jan 29;72(2):197–209. doi: 10.1016/0092-8674(93)90660-i. [DOI] [PubMed] [Google Scholar]
  46. Oppenheim R. W. Cell death during development of the nervous system. Annu Rev Neurosci. 1991;14:453–501. doi: 10.1146/annurev.ne.14.030191.002321. [DOI] [PubMed] [Google Scholar]
  47. Pignatelli M., Cortés-Canteli M., Santos A., Perez-Castillo A. Involvement of the NGFI-A gene in the differentiation of neuroblastoma cells. FEBS Lett. 1999 Nov 12;461(1-2):37–42. doi: 10.1016/s0014-5793(99)01420-9. [DOI] [PubMed] [Google Scholar]
  48. Pipaon C., Santos A., Perez-Castillo A. Thyroid hormone up-regulates NGFI-A gene expression in rat brain during development. J Biol Chem. 1992 Jan 5;267(1):21–23. [PubMed] [Google Scholar]
  49. Raff M. C., Barres B. A., Burne J. F., Coles H. S., Ishizaki Y., Jacobson M. D. Programmed cell death and the control of cell survival: lessons from the nervous system. Science. 1993 Oct 29;262(5134):695–700. doi: 10.1126/science.8235590. [DOI] [PubMed] [Google Scholar]
  50. Saffen D. W., Cole A. J., Worley P. F., Christy B. A., Ryder K., Baraban J. M. Convulsant-induced increase in transcription factor messenger RNAs in rat brain. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7795–7799. doi: 10.1073/pnas.85.20.7795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Shaul Y. c-Abl: activation and nuclear targets. Cell Death Differ. 2000 Jan;7(1):10–16. doi: 10.1038/sj.cdd.4400626. [DOI] [PubMed] [Google Scholar]
  52. Stiewe T., Pützer B. M. Role of the p53-homologue p73 in E2F1-induced apoptosis. Nat Genet. 2000 Dec;26(4):464–469. doi: 10.1038/82617. [DOI] [PubMed] [Google Scholar]
  53. Sukhatme V. P., Cao X. M., Chang L. C., Tsai-Morris C. H., Stamenkovich D., Ferreira P. C., Cohen D. R., Edwards S. A., Shows T. B., Curran T. A zinc finger-encoding gene coregulated with c-fos during growth and differentiation, and after cellular depolarization. Cell. 1988 Apr 8;53(1):37–43. doi: 10.1016/0092-8674(88)90485-0. [DOI] [PubMed] [Google Scholar]
  54. Temple S., Raff M. C. Clonal analysis of oligodendrocyte development in culture: evidence for a developmental clock that counts cell divisions. Cell. 1986 Mar 14;44(5):773–779. doi: 10.1016/0092-8674(86)90843-3. [DOI] [PubMed] [Google Scholar]
  55. Thiel G., Schoch S., Petersohn D. Regulation of synapsin I gene expression by the zinc finger transcription factor zif268/egr-1. J Biol Chem. 1994 May 27;269(21):15294–15301. [PubMed] [Google Scholar]
  56. Thiel Gerald, Cibelli Giuseppe. Regulation of life and death by the zinc finger transcription factor Egr-1. J Cell Physiol. 2002 Dec;193(3):287–292. doi: 10.1002/jcp.10178. [DOI] [PubMed] [Google Scholar]
  57. Vantini G., Skaper S. D. Neurotrophic factors: from physiology to pharmacology? Pharmacol Res. 1992 Jul-Aug;26(1):1–15. doi: 10.1016/1043-6618(92)90701-c. [DOI] [PubMed] [Google Scholar]
  58. Wang Z. Y., Deuel T. F. An S1 nuclease-sensitive homopurine/homopyrimidine domain in the PDGF A-chain promoter contains a novel binding site for the growth factor-inducible protein EGR-1. Biochem Biophys Res Commun. 1992 Oct 15;188(1):433–439. doi: 10.1016/0006-291x(92)92403-k. [DOI] [PubMed] [Google Scholar]
  59. Watson M. A., Milbrandt J. Expression of the nerve growth factor-regulated NGFI-A and NGFI-B genes in the developing rat. Development. 1990 Sep;110(1):173–183. doi: 10.1242/dev.110.1.173. [DOI] [PubMed] [Google Scholar]
  60. White E. Life, death, and the pursuit of apoptosis. Genes Dev. 1996 Jan 1;10(1):1–15. doi: 10.1101/gad.10.1.1. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES