Skip to main content
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1992 Apr;12(4):1490–1499. doi: 10.1128/mcb.12.4.1490

Transcriptional activation of the CEF-4/9E3 cytokine gene by pp60v-src.

M Dehbi 1, A Mbiguino 1, M Beauchemin 1, G Chatelain 1, P A Bédard 1
PMCID: PMC369590  PMID: 1549106

Abstract

The CEF-4/9E3 gene is expressed constitutively in Rous sarcoma virus (RSV)-transformed cells. This expression is largely determined by an increase in transcription of the gene. In this report, we characterize the regulatory elements responsible for the transformation-dependent activation of CEF-4/9E3. Three sequences corresponding to AP-1, PRD II/kappa B, and TAACGCAATT are involved in the process and therefore define the src-responsive unit (SRU) of the CEF-4 promoter. In constructs containing a deletion of the SRU, multiple copies of AP-1 or PRD II/kappa B, but not TAACGCAATT, led to activation of the promoter. Thus, factors interacting with these elements are constitutively activated in RSV-transformed chicken embryo fibroblasts. In agreement with the results of transient expression assays, protein binding to AP-1, PRD II/kappa B, and TAACGCAATT were more abundant in the nuclei of transformed cells. The expression of the CEF-4 promoter was investigated in cells infected by a temperature-sensitive mutant of RSV. No significant increase in CEF-4 promoter activity was detected early after activation of pp60v-src. In contrast, a substantial activation of the CEF-4 promoter was detected late after a temperature shift. Factors interacting with the TAACGCAATT, PRD II/kappa B, and AP-1 elements accumulated gradually over a period of several hours. Therefore, transcriptional activation plays an important role in the late, constitutive expression of the CEF-4 gene in stably transformed cells.

Full text

PDF
1490

Images in this article

Selected References

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

  1. Adkins B., Leutz A., Graf T. Autocrine growth induced by src-related oncogenes in transformed chicken myeloid cells. Cell. 1984 Dec;39(3 Pt 2):439–445. doi: 10.1016/0092-8674(84)90451-3. [DOI] [PubMed] [Google Scholar]
  2. Almendral J. M., Sommer D., Macdonald-Bravo H., Burckhardt J., Perera J., Bravo R. Complexity of the early genetic response to growth factors in mouse fibroblasts. Mol Cell Biol. 1988 May;8(5):2140–2148. doi: 10.1128/mcb.8.5.2140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Anisowicz A., Bardwell L., Sager R. Constitutive overexpression of a growth-regulated gene in transformed Chinese hamster and human cells. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7188–7192. doi: 10.1073/pnas.84.20.7188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Anzano M. A., Roberts A. B., Smith J. M., Sporn M. B., De Larco J. E. Sarcoma growth factor from conditioned medium of virally transformed cells is composed of both type alpha and type beta transforming growth factors. Proc Natl Acad Sci U S A. 1983 Oct;80(20):6264–6268. doi: 10.1073/pnas.80.20.6264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Baichwal V. R., Park A., Tjian R. v-Src and EJ Ras alleviate repression of c-Jun by a cell-specific inhibitor. Nature. 1991 Jul 11;352(6331):165–168. doi: 10.1038/352165a0. [DOI] [PubMed] [Google Scholar]
  6. Barker K., Aderem A., Hanafusa H. Modulation of arachidonic acid metabolism by Rous sarcoma virus. J Virol. 1989 Jul;63(7):2929–2935. doi: 10.1128/jvi.63.7.2929-2935.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Barker K., Hanafusa H. Expression of 9E3 mRNA is associated with mitogenicity, phosphorylation, and morphological alteration in chicken embryo fibroblasts. Mol Cell Biol. 1990 Jul;10(7):3813–3817. doi: 10.1128/mcb.10.7.3813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bedard P. A., Alcorta D., Simmons D. L., Luk K. C., Erikson R. L. Constitutive expression of a gene encoding a polypeptide homologous to biologically active human platelet protein in Rous sarcoma virus-transformed fibroblasts. Proc Natl Acad Sci U S A. 1987 Oct;84(19):6715–6719. doi: 10.1073/pnas.84.19.6715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Bell S. M., Brackenbury R. W., Leslie N. D., Degen J. L. Plasminogen activator gene expression is induced by the src oncogene product and tumor promoters. J Biol Chem. 1990 Jan 25;265(3):1333–1338. [PubMed] [Google Scholar]
  10. Binétruy B., Smeal T., Karin M. Ha-Ras augments c-Jun activity and stimulates phosphorylation of its activation domain. Nature. 1991 May 9;351(6322):122–127. doi: 10.1038/351122a0. [DOI] [PubMed] [Google Scholar]
  11. Birchenall-Roberts M. C., Ruscetti F. W., Kasper J., Lee H. D., Friedman R., Geiser A., Sporn M. B., Roberts A. B., Kim S. J. Transcriptional regulation of the transforming growth factor beta 1 promoter by v-src gene products is mediated through the AP-1 complex. Mol Cell Biol. 1990 Sep;10(9):4978–4983. doi: 10.1128/mcb.10.9.4978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Birnbaum M. J., Haspel H. C., Rosen O. M. Transformation of rat fibroblasts by FSV rapidly increases glucose transporter gene transcription. Science. 1987 Mar 20;235(4795):1495–1498. doi: 10.1126/science.3029870. [DOI] [PubMed] [Google Scholar]
  13. Blobel G. A., Hanafusa H. The v-src inducible gene 9E3/pCEF4 is regulated by both its promoter upstream sequence and its 3' untranslated region. Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1162–1166. doi: 10.1073/pnas.88.4.1162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Bohmann D., Admon A., Turner D. R., Tjian R. Transcriptional regulation by the AP-1 family of enhancer-binding proteins: a nuclear target for signal transduction. Cold Spring Harb Symp Quant Biol. 1988;53(Pt 2):695–700. doi: 10.1101/sqb.1988.053.01.079. [DOI] [PubMed] [Google Scholar]
  15. Bowen-Pope D. F., Vogel A., Ross R. Production of platelet-derived growth factor-like molecules and reduced expression of platelet-derived growth factor receptors accompany transformation by a wide spectrum of agents. Proc Natl Acad Sci U S A. 1984 Apr;81(8):2396–2400. doi: 10.1073/pnas.81.8.2396. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Boyle W. J., Smeal T., Defize L. H., Angel P., Woodgett J. R., Karin M., Hunter T. Activation of protein kinase C decreases phosphorylation of c-Jun at sites that negatively regulate its DNA-binding activity. Cell. 1991 Feb 8;64(3):573–584. doi: 10.1016/0092-8674(91)90241-p. [DOI] [PubMed] [Google Scholar]
  17. Caput D., Beutler B., Hartog K., Thayer R., Brown-Shimer S., Cerami A. Identification of a common nucleotide sequence in the 3'-untranslated region of mRNA molecules specifying inflammatory mediators. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1670–1674. doi: 10.1073/pnas.83.6.1670. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Cochran B. H., Reffel A. C., Stiles C. D. Molecular cloning of gene sequences regulated by platelet-derived growth factor. Cell. 1983 Jul;33(3):939–947. doi: 10.1016/0092-8674(83)90037-5. [DOI] [PubMed] [Google Scholar]
  19. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Dodgson J. B., Strommer J., Engel J. D. Isolation of the chicken beta-globin gene and a linked embryonic beta-like globin gene from a chicken DNA recombinant library. Cell. 1979 Aug;17(4):879–887. doi: 10.1016/0092-8674(79)90328-3. [DOI] [PubMed] [Google Scholar]
  21. Dutta A., Hamaguchi M., Hanafusa H. Serum independence of transcription from the promoter of an avian retrovirus in v-src-transformed cells is a primary, intracellular effect of increased tyrosine phosphorylation. Proc Natl Acad Sci U S A. 1990 Jan;87(2):608–612. doi: 10.1073/pnas.87.2.608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Dutta A., Stoeckle M. Y., Hanafusa H. Serum and v-src increase the level of a CCAAT-binding factor required for transcription from a retroviral long terminal repeat. Genes Dev. 1990 Feb;4(2):243–254. doi: 10.1101/gad.4.2.243. [DOI] [PubMed] [Google Scholar]
  23. Edlund T., Walker M. D., Barr P. J., Rutter W. J. Cell-specific expression of the rat insulin gene: evidence for role of two distinct 5' flanking elements. Science. 1985 Nov 22;230(4728):912–916. doi: 10.1126/science.3904002. [DOI] [PubMed] [Google Scholar]
  24. Flier J. S., Mueckler M. M., Usher P., Lodish H. F. Elevated levels of glucose transport and transporter messenger RNA are induced by ras or src oncogenes. Science. 1987 Mar 20;235(4795):1492–1495. doi: 10.1126/science.3103217. [DOI] [PubMed] [Google Scholar]
  25. Fromental C., Kanno M., Nomiyama H., Chambon P. Cooperativity and hierarchical levels of functional organization in the SV40 enhancer. Cell. 1988 Sep 23;54(7):943–953. doi: 10.1016/0092-8674(88)90109-2. [DOI] [PubMed] [Google Scholar]
  26. Fujii M., Shalloway D., Verma I. M. Gene regulation by tyrosine kinases: src protein activates various promoters, including c-fos. Mol Cell Biol. 1989 Jun;9(6):2493–2499. doi: 10.1128/mcb.9.6.2493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Gauthier-Rouvière C., Fernandez A., Lamb N. J. ras-induced c-fos expression and proliferation in living rat fibroblasts involves C-kinase activation and the serum response element pathway. EMBO J. 1990 Jan;9(1):171–180. doi: 10.1002/j.1460-2075.1990.tb08093.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Gonneville L., Martins T. J., Bédard P. A. Complex expression pattern of the CEF-4 cytokine in transformed and mitogenically stimulated cells. Oncogene. 1991 Oct;6(10):1825–1833. [PubMed] [Google Scholar]
  29. Goodbourn S., Maniatis T. Overlapping positive and negative regulatory domains of the human beta-interferon gene. Proc Natl Acad Sci U S A. 1988 Mar;85(5):1447–1451. doi: 10.1073/pnas.85.5.1447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Greenberg M. E., Ziff E. B. Stimulation of 3T3 cells induces transcription of the c-fos proto-oncogene. Nature. 1984 Oct 4;311(5985):433–438. doi: 10.1038/311433a0. [DOI] [PubMed] [Google Scholar]
  31. Gutman A., Wasylyk B. Nuclear targets for transcription regulation by oncogenes. Trends Genet. 1991 Feb;7(2):49–54. doi: 10.1016/0168-9525(91)90231-E. [DOI] [PubMed] [Google Scholar]
  32. Gutman A., Wasylyk B. The collagenase gene promoter contains a TPA and oncogene-responsive unit encompassing the PEA3 and AP-1 binding sites. EMBO J. 1990 Jul;9(7):2241–2246. doi: 10.1002/j.1460-2075.1990.tb07394.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Gutman A., Wasylyk C., Wasylyk B. Cell-specific regulation of oncogene-responsive sequences of the c-fos promoter. Mol Cell Biol. 1991 Oct;11(10):5381–5387. doi: 10.1128/mcb.11.10.5381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Jamal S., Ziff E. Transactivation of c-fos and beta-actin genes by raf as a step in early response to transmembrane signals. Nature. 1990 Mar 29;344(6265):463–466. doi: 10.1038/344463a0. [DOI] [PubMed] [Google Scholar]
  35. Jähner D., Hunter T. The ras-related gene rhoB is an immediate-early gene inducible by v-Fps, epidermal growth factor, and platelet-derived growth factor in rat fibroblasts. Mol Cell Biol. 1991 Jul;11(7):3682–3690. doi: 10.1128/mcb.11.7.3682. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Jähner D., Hunter T. The stimulation of quiescent rat fibroblasts by v-src and v-fps oncogenic protein-tyrosine kinases leads to the induction of a subset of immediate early genes. Oncogene. 1991 Jul;6(7):1259–1268. [PubMed] [Google Scholar]
  37. Kowalski J., Denhardt D. T. Regulation of the mRNA for monocyte-derived neutrophil-activating peptide in differentiating HL60 promyelocytes. Mol Cell Biol. 1989 May;9(5):1946–1957. doi: 10.1128/mcb.9.5.1946. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Lau L. F., Nathans D. Expression of a set of growth-related immediate early genes in BALB/c 3T3 cells: coordinate regulation with c-fos or c-myc. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1182–1186. doi: 10.1073/pnas.84.5.1182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Lenardo M. J., Fan C. M., Maniatis T., Baltimore D. The involvement of NF-kappa B in beta-interferon gene regulation reveals its role as widely inducible mediator of signal transduction. Cell. 1989 Apr 21;57(2):287–294. doi: 10.1016/0092-8674(89)90966-5. [DOI] [PubMed] [Google Scholar]
  40. Lloyd A., Yancheva N., Wasylyk B. Transformation suppressor activity of a Jun transcription factor lacking its activation domain. Nature. 1991 Aug 15;352(6336):635–638. doi: 10.1038/352635a0. [DOI] [PubMed] [Google Scholar]
  41. Lopata M. A., Cleveland D. W., Sollner-Webb B. High level transient expression of a chloramphenicol acetyl transferase gene by DEAE-dextran mediated DNA transfection coupled with a dimethyl sulfoxide or glycerol shock treatment. Nucleic Acids Res. 1984 Jul 25;12(14):5707–5717. doi: 10.1093/nar/12.14.5707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Matrisian L. M., Bowden G. T., Krieg P., Fürstenberger G., Briand J. P., Leroy P., Breathnach R. The mRNA coding for the secreted protease transin is expressed more abundantly in malignant than in benign tumors. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9413–9417. doi: 10.1073/pnas.83.24.9413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Matrisian L. M., Glaichenhaus N., Gesnel M. C., Breathnach R. Epidermal growth factor and oncogenes induce transcription of the same cellular mRNA in rat fibroblasts. EMBO J. 1985 Jun;4(6):1435–1440. doi: 10.1002/j.1460-2075.1985.tb03799.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. McKnight S., Tjian R. Transcriptional selectivity of viral genes in mammalian cells. Cell. 1986 Sep 12;46(6):795–805. doi: 10.1016/0092-8674(86)90061-9. [DOI] [PubMed] [Google Scholar]
  45. Piette J., Hirai S., Yaniv M. Constitutive synthesis of activator protein 1 transcription factor after viral transformation of mouse fibroblasts. Proc Natl Acad Sci U S A. 1988 May;85(10):3401–3405. doi: 10.1073/pnas.85.10.3401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Qureshi S. A., Cao X. M., Sukhatme V. P., Foster D. A. v-Src activates mitogen-responsive transcription factor Egr-1 via serum response elements. J Biol Chem. 1991 Jun 15;266(17):10802–10806. [PubMed] [Google Scholar]
  47. Qureshi S. A., Joseph C. K., Rim M., Maroney A., Foster D. A. v-Src activates both protein kinase C-dependent and independent signaling pathways in murine fibroblasts. Oncogene. 1991 Jun;6(6):995–999. [PubMed] [Google Scholar]
  48. Richmond A., Thomas H. G. Purification of melanoma growth stimulatory activity. J Cell Physiol. 1986 Dec;129(3):375–384. doi: 10.1002/jcp.1041290316. [DOI] [PubMed] [Google Scholar]
  49. Rosenthal N. Identification of regulatory elements of cloned genes with functional assays. Methods Enzymol. 1987;152:704–720. doi: 10.1016/0076-6879(87)52075-4. [DOI] [PubMed] [Google Scholar]
  50. Salier J. P., Kurachi K. A CAT expression vector with virtually no background: pUMSVOCAT. Biotechniques. 1989 Jan;7(1):30–31. [PubMed] [Google Scholar]
  51. Schönthal A., Herrlich P., Rahmsdorf H. J., Ponta H. Requirement for fos gene expression in the transcriptional activation of collagenase by other oncogenes and phorbol esters. Cell. 1988 Jul 29;54(3):325–334. doi: 10.1016/0092-8674(88)90195-x. [DOI] [PubMed] [Google Scholar]
  52. Shaw G., Kamen R. A conserved AU sequence from the 3' untranslated region of GM-CSF mRNA mediates selective mRNA degradation. Cell. 1986 Aug 29;46(5):659–667. doi: 10.1016/0092-8674(86)90341-7. [DOI] [PubMed] [Google Scholar]
  53. Smeal T., Binetruy B., Mercola D. A., Birrer M., Karin M. Oncogenic and transcriptional cooperation with Ha-Ras requires phosphorylation of c-Jun on serines 63 and 73. Nature. 1991 Dec 12;354(6353):494–496. doi: 10.1038/354494a0. [DOI] [PubMed] [Google Scholar]
  54. Spangler R., Joseph C., Qureshi S. A., Berg K. L., Foster D. A. Evidence that v-src and v-fps gene products use a protein kinase C-mediated pathway to induce expression of a transformation-related gene. Proc Natl Acad Sci U S A. 1989 Sep;86(18):7017–7021. doi: 10.1073/pnas.86.18.7017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Stoeckle M. Y., Barker K. A. Two burgeoning families of platelet factor 4-related proteins: mediators of the inflammatory response. New Biol. 1990 Apr;2(4):313–323. [PubMed] [Google Scholar]
  56. Stoeckle M. Y., Hanafusa H. Processing of 9E3 mRNA and regulation of its stability in normal and Rous sarcoma virus-transformed cells. Mol Cell Biol. 1989 Nov;9(11):4738–4745. doi: 10.1128/mcb.9.11.4738. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Stoeckle M. Y., Sugano S., Hampe A., Vashistha A., Pellman D., Hanafusa H. 78-kilodalton glucose-regulated protein is induced in Rous sarcoma virus-transformed cells independently of glucose deprivation. Mol Cell Biol. 1988 Jul;8(7):2675–2680. doi: 10.1128/mcb.8.7.2675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Sugano S., Stoeckle M. Y., Hanafusa H. Transformation by Rous sarcoma virus induces a novel gene with homology to a mitogenic platelet protein. Cell. 1987 May 8;49(3):321–328. doi: 10.1016/0092-8674(87)90284-4. [DOI] [PubMed] [Google Scholar]
  59. Taylor J. W., Ott J., Eckstein F. The rapid generation of oligonucleotide-directed mutations at high frequency using phosphorothioate-modified DNA. Nucleic Acids Res. 1985 Dec 20;13(24):8765–8785. doi: 10.1093/nar/13.24.8765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Wasylyk C., Gutman A., Nicholson R., Wasylyk B. The c-Ets oncoprotein activates the stromelysin promoter through the same elements as several non-nuclear oncoproteins. EMBO J. 1991 May;10(5):1127–1134. doi: 10.1002/j.1460-2075.1991.tb08053.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Wasylyk C., Imler J. L., Wasylyk B. Transforming but not immortalizing oncogenes activate the transcription factor PEA1. EMBO J. 1988 Aug;7(8):2475–2483. doi: 10.1002/j.1460-2075.1988.tb03094.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Welham M. J., Wyke J. A., Lang A., Wyke A. W. Mitogenesis induced by pp60v-src is not accompanied by increased expression of immediate early response genes. Oncogene. 1990 Feb;5(2):161–169. [PubMed] [Google Scholar]
  63. White M. K., Rall T. B., Weber M. J. Differential regulation of glucose transporter isoforms by the src oncogene in chicken embryo fibroblasts. Mol Cell Biol. 1991 Sep;11(9):4448–4454. doi: 10.1128/mcb.11.9.4448. [DOI] [PMC free article] [PubMed] [Google Scholar]
  64. Wood T. G., McGeady M. L., Baroudy B. M., Blair D. G., Vande Woude G. F. Mouse c-mos oncogene activation is prevented by upstream sequences. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7817–7821. doi: 10.1073/pnas.81.24.7817. [DOI] [PMC free article] [PubMed] [Google Scholar]
  65. Yamamura Y., Hattori T., Obaru K., Sakai K., Asou N., Takatsuki K., Ohmoto Y., Nomiyama H., Shimada K. Synthesis of a novel cytokine and its gene (LD78) expressions in hematopoietic fresh tumor cells and cell lines. J Clin Invest. 1989 Dec;84(6):1707–1712. doi: 10.1172/JCI114353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  66. Zipfel P. F., Balke J., Irving S. G., Kelly K., Siebenlist U. Mitogenic activation of human T cells induces two closely related genes which share structural similarities with a new family of secreted factors. J Immunol. 1989 Mar 1;142(5):1582–1590. [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES