Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 2002 Dec 1;368(Pt 2):573–580. doi: 10.1042/BJ20020579

Dual role for mitogen-activated protein kinase (Erk) in insulin-dependent regulation of Fra-1 (fos-related antigen-1) transcription and phosphorylation.

Toby W Hurd 1, Ainsley A Culbert 1, Kenneth J Webster 1, Jeremy M Tavaré 1
PMCID: PMC1223008  PMID: 12197835

Abstract

Insulin regulates the activity of the AP-1 (activator protein-1) transcriptional complex in several cell types. One component of the AP-1 complex is the transcription factor Fra-1 (fos-related antigen-1), and we have demonstrated previously that insulin stimulates the expression of Fra-1 mRNA in CHO.T cells [Griffiths, Black, Culbert, Dickens, Shaw, Gillespie and Tavaré (1998) Biochem. J. 335, 19-26]. Here we demonstrate that insulin stimulates the activity of a fra-1 promoter linked to a luciferase reporter gene, indicating that the ability of insulin to induce expression of Fra-1 mRNA is due, at least in part, to an increase in gene transcription. Furthermore, we found that insulin induces the serine phosphorylation of Fra-1 and reduces its mobility during SDS/PAGE as a result of phosphorylation. The ability of insulin to induce the accumulation of Fra-1 mRNA, stimulate the fra-1 promoter and stimulate phosphorylation of Fra-1 all require the mitogen-activated protein (MAP) kinase cascade, which leads to the activation of extracellular-signal-regulated kinase (Erk) 1/2. Consequently, our results demonstrate that the Erk cascade plays a dual role in the co-ordinated regulation of the transcription and the phosphorylation of Fra-1 by insulin.

Full Text

The Full Text of this article is available as a PDF (213.6 KB).

Selected References

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

  1. 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]
  2. Bergers G., Graninger P., Braselmann S., Wrighton C., Busslinger M. Transcriptional activation of the fra-1 gene by AP-1 is mediated by regulatory sequences in the first intron. Mol Cell Biol. 1995 Jul;15(7):3748–3758. doi: 10.1128/mcb.15.7.3748. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cook S. J., Aziz N., McMahon M. The repertoire of fos and jun proteins expressed during the G1 phase of the cell cycle is determined by the duration of mitogen-activated protein kinase activation. Mol Cell Biol. 1999 Jan;19(1):330–341. doi: 10.1128/mcb.19.1.330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ellis L., Clauser E., Morgan D. O., Edery M., Roth R. A., Rutter W. J. Replacement of insulin receptor tyrosine residues 1162 and 1163 compromises insulin-stimulated kinase activity and uptake of 2-deoxyglucose. Cell. 1986 Jun 6;45(5):721–732. doi: 10.1016/0092-8674(86)90786-5. [DOI] [PubMed] [Google Scholar]
  5. Graham F. L., Prevec L. Methods for construction of adenovirus vectors. Mol Biotechnol. 1995 Jun;3(3):207–220. doi: 10.1007/BF02789331. [DOI] [PubMed] [Google Scholar]
  6. Graham F. L., Smiley J., Russell W. C., Nairn R. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol. 1977 Jul;36(1):59–74. doi: 10.1099/0022-1317-36-1-59. [DOI] [PubMed] [Google Scholar]
  7. Griffiths M. R., Black E. J., Culbert A. A., Dickens M., Shaw P. E., Gillespie D. A., Tavaré J. M. Insulin-stimulated expression of c-fos, fra1 and c-jun accompanies the activation of the activator protein-1 (AP-1) transcriptional complex. Biochem J. 1998 Oct 1;335(Pt 1):19–26. doi: 10.1042/bj3350019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Grigoriadis A. E., Schellander K., Wang Z. Q., Wagner E. F. Osteoblasts are target cells for transformation in c-fos transgenic mice. J Cell Biol. 1993 Aug;122(3):685–701. doi: 10.1083/jcb.122.3.685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gruda M. C., Kovary K., Metz R., Bravo R. Regulation of Fra-1 and Fra-2 phosphorylation differs during the cell cycle of fibroblasts and phosphorylation in vitro by MAP kinase affects DNA binding activity. Oncogene. 1994 Sep;9(9):2537–2547. [PubMed] [Google Scholar]
  10. Harding T. C., Geddes B. J., Noel J. D., Murphy D., Uney J. B. Tetracycline-regulated transgene expression in hippocampal neurones following transfection with adenoviral vectors. J Neurochem. 1997 Dec;69(6):2620–2623. doi: 10.1046/j.1471-4159.1997.69062620.x. [DOI] [PubMed] [Google Scholar]
  11. Hill C. S., Treisman R. Transcriptional regulation by extracellular signals: mechanisms and specificity. Cell. 1995 Jan 27;80(2):199–211. doi: 10.1016/0092-8674(95)90403-4. [DOI] [PubMed] [Google Scholar]
  12. McGrory W. J., Bautista D. S., Graham F. L. A simple technique for the rescue of early region I mutations into infectious human adenovirus type 5. Virology. 1988 Apr;163(2):614–617. doi: 10.1016/0042-6822(88)90302-9. [DOI] [PubMed] [Google Scholar]
  13. O'Brien R. M., Granner D. K. Regulation of gene expression by insulin. Physiol Rev. 1996 Oct;76(4):1109–1161. doi: 10.1152/physrev.1996.76.4.1109. [DOI] [PubMed] [Google Scholar]
  14. Rutter G. A., White M. R., Tavaré J. M. Involvement of MAP kinase in insulin signalling revealed by non-invasive imaging of luciferase gene expression in single living cells. Curr Biol. 1995 Aug 1;5(8):890–899. doi: 10.1016/s0960-9822(95)00179-5. [DOI] [PubMed] [Google Scholar]
  15. Schaeffer H. J., Catling A. D., Eblen S. T., Collier L. S., Krauss A., Weber M. J. MP1: a MEK binding partner that enhances enzymatic activation of the MAP kinase cascade. Science. 1998 Sep 11;281(5383):1668–1671. doi: 10.1126/science.281.5383.1668. [DOI] [PubMed] [Google Scholar]
  16. Schreiber M., Wang Z. Q., Jochum W., Fetka I., Elliott C., Wagner E. F. Placental vascularisation requires the AP-1 component fra1. Development. 2000 Nov;127(22):4937–4948. doi: 10.1242/dev.127.22.4937. [DOI] [PubMed] [Google Scholar]
  17. Sonobe M. H., Yoshida T., Murakami M., Kameda T., Iba H. fra-2 promoter can respond to serum-stimulation through AP-1 complexes. Oncogene. 1995 Feb 16;10(4):689–696. [PubMed] [Google Scholar]
  18. Streeper R. S., Chapman S. C., Ayala J. E., Svitek C. A., Goldman J. K., Cave A., O'Brien R. M. A phorbol ester-insensitive AP-1 motif mediates the stimulatory effect of insulin on rat malic enzyme gene transcription. Mol Endocrinol. 1998 Nov;12(11):1778–1791. doi: 10.1210/mend.12.11.0199. [DOI] [PubMed] [Google Scholar]
  19. Tavaré J. M., Issad T. Two-dimensional phosphopeptide mapping of receptor tyrosine kinases. Methods Mol Biol. 2001;124:67–85. doi: 10.1385/1-59259-059-4:67. [DOI] [PubMed] [Google Scholar]
  20. Tavaré J. M., O'Brien R. M., Siddle K., Denton R. M. Analysis of insulin-receptor phosphorylation sites in intact cells by two-dimensional phosphopeptide mapping. Biochem J. 1988 Aug 1;253(3):783–788. doi: 10.1042/bj2530783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Thompson M. J., Roe M. W., Malik R. K., Blackshear P. J. Insulin and other growth factors induce binding of the ternary complex and a novel protein complex to the c-fos serum response element. J Biol Chem. 1994 Aug 19;269(33):21127–21135. [PubMed] [Google Scholar]
  22. Treinies I., Paterson H. F., Hooper S., Wilson R., Marshall C. J. Activated MEK stimulates expression of AP-1 components independently of phosphatidylinositol 3-kinase (PI3-kinase) but requires a PI3-kinase signal To stimulate DNA synthesis. Mol Cell Biol. 1999 Jan;19(1):321–329. doi: 10.1128/mcb.19.1.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wisdom R., Verma I. M. Proto-oncogene FosB: the amino terminus encodes a regulatory function required for transformation. Mol Cell Biol. 1993 May;13(5):2635–2643. doi: 10.1128/mcb.13.5.2635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wisdom R., Yen J., Rashid D., Verma I. M. Transformation by FosB requires a trans-activation domain missing in FosB2 that can be substituted by heterologous activation domains. Genes Dev. 1992 Apr;6(4):667–675. doi: 10.1101/gad.6.4.667. [DOI] [PubMed] [Google Scholar]
  25. Wisdon R., Verma I. M. Transformation by Fos proteins requires a C-terminal transactivation domain. Mol Cell Biol. 1993 Dec;13(12):7429–7438. doi: 10.1128/mcb.13.12.7429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Yoshioka K., Deng T., Cavigelli M., Karin M. Antitumor promotion by phenolic antioxidants: inhibition of AP-1 activity through induction of Fra expression. Proc Natl Acad Sci U S A. 1995 May 23;92(11):4972–4976. doi: 10.1073/pnas.92.11.4972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Young Matthew R., Nair Rajalakshmi, Bucheimer Natalie, Tulsian Preety, Brown Nicole, Chapp Cristi, Hsu Tin-Chen, Colburn Nancy H. Transactivation of Fra-1 and consequent activation of AP-1 occur extracellular signal-regulated kinase dependently. Mol Cell Biol. 2002 Jan;22(2):587–598. doi: 10.1128/MCB.22.2.587-598.2002. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES