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. 1997 Jun;17(6):3202–3209. doi: 10.1128/mcb.17.6.3202

The c-Jun-induced transformation process involves complex regulation of tenascin-C expression.

A Mettouchi 1, F Cabon 1, N Montreau 1, V Dejong 1, P Vernier 1, R Gherzi 1, G Mercier 1, B Binétruy 1
PMCID: PMC232173  PMID: 9154819

Abstract

In cooperation with an activated ras oncogene, the site-dependent AP-1 transcription factor c-Jun transforms primary rat embryo fibroblasts (REF). Although signal transduction pathways leading to activation of c-Jun proteins have been extensively studied, little is known about c-Jun cellular targets. We identified c-Jun-upregulated cDNA clones homologous to the tenascin-C gene by differential screening of a cDNA library from REF. This tightly regulated gene encodes a rare extracellular matrix protein involved in cell attachment and migration and in the control of cell growth. Transient overexpression of c-Jun induced tenascin-C expression in primary REF and in FR3T3, an established fibroblast cell line. Surprisingly, tenascin-C synthesis was repressed after stable transformation by c-Jun compared to that in the nontransformed parental cells. As assessed by using the tenascin-C (-220 to +79) promoter fragment cloned in a reporter construct, the c-Jun-induced transient activation is mediated by two binding sites: one GCN4/AP-1-like site, at position -146, and one NF-kappaB site, at position -210. Furthermore, as demonstrated by gel shift experiments and cotransfections of the reporter plasmid and expression vectors encoding the p65 subunit of NF-kappaB and c-Jun, the two transcription factors bind and synergistically transactivate the tenascin-C promoter. We previously described two other extracellular matrix proteins, SPARC and thrombospondin-1, as c-Jun targets. Thus, our results strongly suggest that the regulation of the extracellular matrix composition plays a central role in c-Jun-induced transformation.

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Selected References

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  1. Alani R., Brown P., Binétruy B., Dosaka H., Rosenberg R. K., Angel P., Karin M., Birrer M. J. The transactivating domain of the c-Jun proto-oncoprotein is required for cotransformation of rat embryo cells. Mol Cell Biol. 1991 Dec;11(12):6286–6295. doi: 10.1128/mcb.11.12.6286. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Angel P., Baumann I., Stein B., Delius H., Rahmsdorf H. J., Herrlich P. 12-O-tetradecanoyl-phorbol-13-acetate induction of the human collagenase gene is mediated by an inducible enhancer element located in the 5'-flanking region. Mol Cell Biol. 1987 Jun;7(6):2256–2266. doi: 10.1128/mcb.7.6.2256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Angel P., Imagawa M., Chiu R., Stein B., Imbra R. J., Rahmsdorf H. J., Jonat C., Herrlich P., Karin M. Phorbol ester-inducible genes contain a common cis element recognized by a TPA-modulated trans-acting factor. Cell. 1987 Jun 19;49(6):729–739. doi: 10.1016/0092-8674(87)90611-8. [DOI] [PubMed] [Google Scholar]
  4. Angel P., Karin M. The role of Jun, Fos and the AP-1 complex in cell-proliferation and transformation. Biochim Biophys Acta. 1991 Dec 10;1072(2-3):129–157. doi: 10.1016/0304-419x(91)90011-9. [DOI] [PubMed] [Google Scholar]
  5. Arndt K., Fink G. R. GCN4 protein, a positive transcription factor in yeast, binds general control promoters at all 5' TGACTC 3' sequences. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8516–8520. doi: 10.1073/pnas.83.22.8516. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Aukhil I., Joshi P., Yan Y., Erickson H. P. Cell- and heparin-binding domains of the hexabrachion arm identified by tenascin expression proteins. J Biol Chem. 1993 Feb 5;268(4):2542–2553. [PubMed] [Google Scholar]
  7. Bengal E., Ransone L., Scharfmann R., Dwarki V. J., Tapscott S. J., Weintraub H., Verma I. M. Functional antagonism between c-Jun and MyoD proteins: a direct physical association. Cell. 1992 Feb 7;68(3):507–519. doi: 10.1016/0092-8674(92)90187-h. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Bohmann D., Bos T. J., Admon A., Nishimura T., Vogt P. K., Tjian R. Human proto-oncogene c-jun encodes a DNA binding protein with structural and functional properties of transcription factor AP-1. Science. 1987 Dec 4;238(4832):1386–1392. doi: 10.1126/science.2825349. [DOI] [PubMed] [Google Scholar]
  10. Borsi L., Carnemolla B., Nicolò G., Spina B., Tanara G., Zardi L. Expression of different tenascin isoforms in normal, hyperplastic and neoplastic human breast tissues. Int J Cancer. 1992 Nov 11;52(5):688–692. doi: 10.1002/ijc.2910520504. [DOI] [PubMed] [Google Scholar]
  11. Bos T. J., Monteclaro F. S., Mitsunobu F., Ball A. R., Jr, Chang C. H., Nishimura T., Vogt P. K. Efficient transformation of chicken embryo fibroblasts by c-Jun requires structural modification in coding and noncoding sequences. Genes Dev. 1990 Oct;4(10):1677–1687. doi: 10.1101/gad.4.10.1677. [DOI] [PubMed] [Google Scholar]
  12. Bossy-Wetzel E., Bravo R., Hanahan D. Transcription factors junB and c-jun are selectively up-regulated and functionally implicated in fibrosarcoma development. Genes Dev. 1992 Dec;6(12A):2340–2351. doi: 10.1101/gad.6.12a.2340. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Cano E., Hazzalin C. A., Mahadevan L. C. Anisomycin-activated protein kinases p45 and p55 but not mitogen-activated protein kinases ERK-1 and -2 are implicated in the induction of c-fos and c-jun. Mol Cell Biol. 1994 Nov;14(11):7352–7362. doi: 10.1128/mcb.14.11.7352. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Carnemolla B., Borsi L., Bannikov G., Troyanovsky S., Zardi L. Comparison of human tenascin expression in normal, simian-virus-40-transformed and tumor-derived cell lines. Eur J Biochem. 1992 Apr 15;205(2):561–567. doi: 10.1111/j.1432-1033.1992.tb16813.x. [DOI] [PubMed] [Google Scholar]
  16. Castellazzi M., Dangy J. P., Mechta F., Hirai S., Yaniv M., Samarut J., Lassailly A., Brun G. Overexpression of avian or mouse c-jun in primary chick embryo fibroblasts confers a partially transformed phenotype. Oncogene. 1990 Oct;5(10):1541–1547. [PubMed] [Google Scholar]
  17. Chiquet-Ehrismann R., Mackie E. J., Pearson C. A., Sakakura T. Tenascin: an extracellular matrix protein involved in tissue interactions during fetal development and oncogenesis. Cell. 1986 Oct 10;47(1):131–139. doi: 10.1016/0092-8674(86)90374-0. [DOI] [PubMed] [Google Scholar]
  18. Chiu R., Boyle W. J., Meek J., Smeal T., Hunter T., Karin M. The c-Fos protein interacts with c-Jun/AP-1 to stimulate transcription of AP-1 responsive genes. Cell. 1988 Aug 12;54(4):541–552. doi: 10.1016/0092-8674(88)90076-1. [DOI] [PubMed] [Google Scholar]
  19. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  20. Crossin K. L. Cytotactin binding: inhibition of stimulated proliferation and intracellular alkalinization in fibroblasts. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11403–11407. doi: 10.1073/pnas.88.24.11403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Desbois C., Aubert D., Legrand C., Pain B., Samarut J. A novel mechanism of action for v-ErbA: abrogation of the inactivation of transcription factor AP-1 by retinoic acid and thyroid hormone receptors. Cell. 1991 Nov 15;67(4):731–740. doi: 10.1016/0092-8674(91)90068-a. [DOI] [PubMed] [Google Scholar]
  22. End P., Panayotou G., Entwistle A., Waterfield M. D., Chiquet M. Tenascin: a modulator of cell growth. Eur J Biochem. 1992 Nov 1;209(3):1041–1051. doi: 10.1111/j.1432-1033.1992.tb17380.x. [DOI] [PubMed] [Google Scholar]
  23. Erickson H. P., Bourdon M. A. Tenascin: an extracellular matrix protein prominent in specialized embryonic tissues and tumors. Annu Rev Cell Biol. 1989;5:71–92. doi: 10.1146/annurev.cb.05.110189.000443. [DOI] [PubMed] [Google Scholar]
  24. Erickson H. P. Tenascin-C, tenascin-R and tenascin-X: a family of talented proteins in search of functions. Curr Opin Cell Biol. 1993 Oct;5(5):869–876. doi: 10.1016/0955-0674(93)90037-q. [DOI] [PubMed] [Google Scholar]
  25. Gherzi R., Carnemolla B., Siri A., Ponassi M., Balza E., Zardi L. Human tenascin gene. Structure of the 5'-region, identification, and characterization of the transcription regulatory sequences. J Biol Chem. 1995 Feb 17;270(7):3429–3434. doi: 10.1074/jbc.270.7.3429. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Hiraiwa N., Kida H., Sakakura T., Kusakabe M. Induction of tenascin in cancer cells by interactions with embryonic mesenchyme mediated by a diffusible factor. J Cell Sci. 1993 Feb;104(Pt 2):289–296. doi: 10.1242/jcs.104.2.289. [DOI] [PubMed] [Google Scholar]
  28. Husmann K., Faissner A., Schachner M. Tenascin promotes cerebellar granule cell migration and neurite outgrowth by different domains in the fibronectin type III repeats. J Cell Biol. 1992 Mar;116(6):1475–1486. doi: 10.1083/jcb.116.6.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Joshi P., Chung C. Y., Aukhil I., Erickson H. P. Endothelial cells adhere to the RGD domain and the fibrinogen-like terminal knob of tenascin. J Cell Sci. 1993 Sep;106(Pt 1):389–400. doi: 10.1242/jcs.106.1.389. [DOI] [PubMed] [Google Scholar]
  30. Karin M., Hunter T. Transcriptional control by protein phosphorylation: signal transmission from the cell surface to the nucleus. Curr Biol. 1995 Jul 1;5(7):747–757. doi: 10.1016/s0960-9822(95)00151-5. [DOI] [PubMed] [Google Scholar]
  31. Karin M. The regulation of AP-1 activity by mitogen-activated protein kinases. J Biol Chem. 1995 Jul 14;270(28):16483–16486. doi: 10.1074/jbc.270.28.16483. [DOI] [PubMed] [Google Scholar]
  32. Kieran M., Blank V., Logeat F., Vandekerckhove J., Lottspeich F., Le Bail O., Urban M. B., Kourilsky P., Baeuerle P. A., Israël A. The DNA binding subunit of NF-kappa B is identical to factor KBF1 and homologous to the rel oncogene product. Cell. 1990 Sep 7;62(5):1007–1018. doi: 10.1016/0092-8674(90)90275-j. [DOI] [PubMed] [Google Scholar]
  33. Lochter A., Vaughan L., Kaplony A., Prochiantz A., Schachner M., Faissner A. J1/tenascin in substrate-bound and soluble form displays contrary effects on neurite outgrowth. J Cell Biol. 1991 Jun;113(5):1159–1171. doi: 10.1083/jcb.113.5.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Mackie E. J., Chiquet-Ehrismann R., Pearson C. A., Inaguma Y., Taya K., Kawarada Y., Sakakura T. Tenascin is a stromal marker for epithelial malignancy in the mammary gland. Proc Natl Acad Sci U S A. 1987 Jul;84(13):4621–4625. doi: 10.1073/pnas.84.13.4621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Mackie E. J., Halfter W., Liverani D. Induction of tenascin in healing wounds. J Cell Biol. 1988 Dec;107(6 Pt 2):2757–2767. doi: 10.1083/jcb.107.6.2757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Mettouchi A., Cabon F., Montreau N., Vernier P., Mercier G., Blangy D., Tricoire H., Vigier P., Binétruy B. SPARC and thrombospondin genes are repressed by the c-jun oncogene in rat embryo fibroblasts. EMBO J. 1994 Dec 1;13(23):5668–5678. doi: 10.1002/j.1460-2075.1994.tb06905.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Nolan G. P. NF-AT-AP-1 and Rel-bZIP: hybrid vigor and binding under the influence. Cell. 1994 Jun 17;77(6):795–798. doi: 10.1016/0092-8674(94)90126-0. [DOI] [PubMed] [Google Scholar]
  38. Prieto A. L., Andersson-Fisone C., Crossin K. L. Characterization of multiple adhesive and counteradhesive domains in the extracellular matrix protein cytotactin. J Cell Biol. 1992 Nov;119(3):663–678. doi: 10.1083/jcb.119.3.663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Saga Y., Tsukamoto T., Jing N., Kusakabe M., Sakakura T. Murine tenascin: cDNA cloning, structure and temporal expression of isoforms. Gene. 1991 Aug 15;104(2):177–185. doi: 10.1016/0378-1119(91)90248-a. [DOI] [PubMed] [Google Scholar]
  40. Saga Y., Yagi T., Ikawa Y., Sakakura T., Aizawa S. Mice develop normally without tenascin. Genes Dev. 1992 Oct;6(10):1821–1831. doi: 10.1101/gad.6.10.1821. [DOI] [PubMed] [Google Scholar]
  41. Schmitz M. L., Baeuerle P. A. The p65 subunit is responsible for the strong transcription activating potential of NF-kappa B. EMBO J. 1991 Dec;10(12):3805–3817. doi: 10.1002/j.1460-2075.1991.tb04950.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Schütte J., Minna J. D., Birrer M. J. Deregulated expression of human c-jun transforms primary rat embryo cells in cooperation with an activated c-Ha-ras gene and transforms rat-1a cells as a single gene. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2257–2261. doi: 10.1073/pnas.86.7.2257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Sharifi B. G., LaFleur D. W., Pirola C. J., Forrester J. S., Fagin J. A. Angiotensin II regulates tenascin gene expression in vascular smooth muscle cells. J Biol Chem. 1992 Nov 25;267(33):23910–23915. [PubMed] [Google Scholar]
  44. 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]
  45. Stein B., Baldwin A. S., Jr, Ballard D. W., Greene W. C., Angel P., Herrlich P. Cross-coupling of the NF-kappa B p65 and Fos/Jun transcription factors produces potentiated biological function. EMBO J. 1993 Oct;12(10):3879–3891. doi: 10.1002/j.1460-2075.1993.tb06066.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Steindler D. A., Settles D., Erickson H. P., Laywell E. D., Yoshiki A., Faissner A., Kusakabe M. Tenascin knockout mice: barrels, boundary molecules, and glial scars. J Neurosci. 1995 Mar;15(3 Pt 1):1971–1983. doi: 10.1523/JNEUROSCI.15-03-01971.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Struhl K. The DNA-binding domains of the jun oncoprotein and the yeast GCN4 transcriptional activator protein are functionally homologous. Cell. 1987 Sep 11;50(6):841–846. doi: 10.1016/0092-8674(87)90511-3. [DOI] [PubMed] [Google Scholar]
  48. Vandel L., Montreau N., Vial E., Pfarr C. M., Binetruy B., Castellazzi M. Stepwise transformation of rat embryo fibroblasts: c-Jun, JunB, or JunD can cooperate with Ras for focus formation, but a c-Jun-containing heterodimer is required for immortalization. Mol Cell Biol. 1996 May;16(5):1881–1888. doi: 10.1128/mcb.16.5.1881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Vincent S., Marty L., Fort P. S26 ribosomal protein RNA: an invariant control for gene regulation experiments in eucaryotic cells and tissues. Nucleic Acids Res. 1993 Mar 25;21(6):1498–1498. doi: 10.1093/nar/21.6.1498. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Vogt P. K., Bos T. J. jun: oncogene and transcription factor. Adv Cancer Res. 1990;55:1–35. doi: 10.1016/s0065-230x(08)60466-2. [DOI] [PubMed] [Google Scholar]
  51. Wehrle B., Chiquet M. Tenascin is accumulated along developing peripheral nerves and allows neurite outgrowth in vitro. Development. 1990 Oct;110(2):401–415. doi: 10.1242/dev.110.2.401. [DOI] [PubMed] [Google Scholar]
  52. Xia Z., Dickens M., Raingeaud J., Davis R. J., Greenberg M. E. Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. Science. 1995 Nov 24;270(5240):1326–1331. doi: 10.1126/science.270.5240.1326. [DOI] [PubMed] [Google Scholar]
  53. Yang-Yen H. F., Chambard J. C., Sun Y. L., Smeal T., Schmidt T. J., Drouin J., Karin M. Transcriptional interference between c-Jun and the glucocorticoid receptor: mutual inhibition of DNA binding due to direct protein-protein interaction. Cell. 1990 Sep 21;62(6):1205–1215. doi: 10.1016/0092-8674(90)90396-v. [DOI] [PubMed] [Google Scholar]

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