Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1996 Aug 1;134(3):747–755. doi: 10.1083/jcb.134.3.747

Regulation of tyrosinase gene expression by cAMP in B16 melanoma cells involves two CATGTG motifs surrounding the TATA box: implication of the microphthalmia gene product

PMCID: PMC2120943  PMID: 8707852

Abstract

In melanocytes and in melanoma cells, upregulation of melanogenesis, by cAMP elevating agents, results from a stimulation of tyrosinase activity that has been ascribed to an increase in tyrosinase protein and messenger amount. However, the mechanism by which cAMP elevating agents increase tyrosinase mRNA remains to be elucidated. In this study, using a luciferase reporter plasmid containing the 2.2-kb fragment 5' of the transcriptional start site of the mouse tyrosinase gene, we showed that cAMP elevating agents lead to a strong stimulation (20-fold) of transcriptional activity of the tyrosinase promoter. Deletions and mutations in the mouse tyrosinase promoter showed that the M-box 70-bp upstream from the TATA-box and the E-box located downstream the TATA-box, near to the initiator site, are involved in the regulation of the tyrosinase promoter activity by cAMP. Additionally, we showed that microphthalmia, a b-HLH transcription factor associated with pigmentation disorders in mouse, binds to these regulatory elements and modulates the transcriptional activity of the tyrosinase promoter. Since cAMP stimulates the binding of microphthalmia to the M-box and to the E-box; it is tempting to propose that microphthalmia, through its interaction with cis-acting elements surrounding the TATA-box, plays a key role in the regulation of the mouse tyrosinase gene expression by cAMP.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

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

  1. Abdel-Malek Z., Swope V., Collins C., Boissy R., Zhao H., Nordlund J. Contribution of melanogenic proteins to the heterogeneous pigmentation of human melanocytes. J Cell Sci. 1993 Dec;106(Pt 4):1323–1331. doi: 10.1242/jcs.106.4.1323. [DOI] [PubMed] [Google Scholar]
  2. Aberdam E., Roméro C., Ortonne J. P. Repeated UVB irradiations do not have the same potential to promote stimulation of melanogenesis in cultured normal human melanocytes. J Cell Sci. 1993 Dec;106(Pt 4):1015–1022. doi: 10.1242/jcs.106.4.1015. [DOI] [PubMed] [Google Scholar]
  3. Agin P. P., Dowdy J. C., Costlow M. E. Diacylglycerol-induced melanogenesis in Skh-2 pigmented hairless mice. Photodermatol Photoimmunol Photomed. 1991 Apr;8(2):51–56. [PubMed] [Google Scholar]
  4. 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]
  5. Aroca P., Urabe K., Kobayashi T., Tsukamoto K., Hearing V. J. Melanin biosynthesis patterns following hormonal stimulation. J Biol Chem. 1993 Dec 5;268(34):25650–25655. [PubMed] [Google Scholar]
  6. Bentley N. J., Eisen T., Goding C. R. Melanocyte-specific expression of the human tyrosinase promoter: activation by the microphthalmia gene product and role of the initiator. Mol Cell Biol. 1994 Dec;14(12):7996–8006. doi: 10.1128/mcb.14.12.7996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Blackwood E. M., Eisenman R. N. Max: a helix-loop-helix zipper protein that forms a sequence-specific DNA-binding complex with Myc. Science. 1991 Mar 8;251(4998):1211–1217. doi: 10.1126/science.2006410. [DOI] [PubMed] [Google Scholar]
  8. Borrelli E., Montmayeur J. P., Foulkes N. S., Sassone-Corsi P. Signal transduction and gene control: the cAMP pathway. Crit Rev Oncog. 1992;3(4):321–338. [PubMed] [Google Scholar]
  9. Burgering B. M., Pronk G. J., van Weeren P. C., Chardin P., Bos J. L. cAMP antagonizes p21ras-directed activation of extracellular signal-regulated kinase 2 and phosphorylation of mSos nucleotide exchange factor. EMBO J. 1993 Nov;12(11):4211–4220. doi: 10.1002/j.1460-2075.1993.tb06105.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Christoffersen J., Smeland E. B., Stokke T., Taskén K., Andersson K. B., Blomhoff H. K. Retinoblastoma protein is rapidly dephosphorylated by elevated cyclic adenosine monophosphate levels in human B-lymphoid cells. Cancer Res. 1994 Apr 15;54(8):2245–2250. [PubMed] [Google Scholar]
  11. 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]
  12. Edmondson D. G., Olson E. N. Helix-loop-helix proteins as regulators of muscle-specific transcription. J Biol Chem. 1993 Jan 15;268(2):755–758. [PubMed] [Google Scholar]
  13. Englaro W., Rezzonico R., Durand-Clément M., Lallemand D., Ortonne J. P., Ballotti R. Mitogen-activated protein kinase pathway and AP-1 are activated during cAMP-induced melanogenesis in B-16 melanoma cells. J Biol Chem. 1995 Oct 13;270(41):24315–24320. doi: 10.1074/jbc.270.41.24315. [DOI] [PubMed] [Google Scholar]
  14. Friedmann P. S., Gilchrest B. A. Ultraviolet radiation directly induces pigment production by cultured human melanocytes. J Cell Physiol. 1987 Oct;133(1):88–94. doi: 10.1002/jcp.1041330111. [DOI] [PubMed] [Google Scholar]
  15. Fuller B. B., Lunsford J. B., Iman D. S. Alpha-melanocyte-stimulating hormone regulation of tyrosinase in Cloudman S-91 mouse melanoma cell cultures. J Biol Chem. 1987 Mar 25;262(9):4024–4033. [PubMed] [Google Scholar]
  16. Ganss R., Schütz G., Beermann F. The mouse tyrosinase gene. Promoter modulation by positive and negative regulatory elements. J Biol Chem. 1994 Nov 25;269(47):29808–29816. [PubMed] [Google Scholar]
  17. Gregor P. D., Sawadogo M., Roeder R. G. The adenovirus major late transcription factor USF is a member of the helix-loop-helix group of regulatory proteins and binds to DNA as a dimer. Genes Dev. 1990 Oct;4(10):1730–1740. doi: 10.1101/gad.4.10.1730. [DOI] [PubMed] [Google Scholar]
  18. Hearing V. J., Jiménez M. Analysis of mammalian pigmentation at the molecular level. Pigment Cell Res. 1989 Mar-Apr;2(2):75–85. doi: 10.1111/j.1600-0749.1989.tb00166.x. [DOI] [PubMed] [Google Scholar]
  19. Hearing V. J., Jiménez M. Mammalian tyrosinase--the critical regulatory control point in melanocyte pigmentation. Int J Biochem. 1987;19(12):1141–1147. doi: 10.1016/0020-711x(87)90095-4. [DOI] [PubMed] [Google Scholar]
  20. Hearing V. J., Tsukamoto K. Enzymatic control of pigmentation in mammals. FASEB J. 1991 Nov;5(14):2902–2909. [PubMed] [Google Scholar]
  21. Hemesath T. J., Steingrímsson E., McGill G., Hansen M. J., Vaught J., Hodgkinson C. A., Arnheiter H., Copeland N. G., Jenkins N. A., Fisher D. E. microphthalmia, a critical factor in melanocyte development, defines a discrete transcription factor family. Genes Dev. 1994 Nov 15;8(22):2770–2780. doi: 10.1101/gad.8.22.2770. [DOI] [PubMed] [Google Scholar]
  22. Hodgkinson C. A., Moore K. J., Nakayama A., Steingrímsson E., Copeland N. G., Jenkins N. A., Arnheiter H. Mutations at the mouse microphthalmia locus are associated with defects in a gene encoding a novel basic-helix-loop-helix-zipper protein. Cell. 1993 Jul 30;74(2):395–404. doi: 10.1016/0092-8674(93)90429-t. [DOI] [PubMed] [Google Scholar]
  23. Hughes A. E., Newton V. E., Liu X. Z., Read A. P. A gene for Waardenburg syndrome type 2 maps close to the human homologue of the microphthalmia gene at chromosome 3p12-p14.1. Nat Genet. 1994 Aug;7(4):509–512. doi: 10.1038/ng0894-509. [DOI] [PubMed] [Google Scholar]
  24. Hughes M. J., Lingrel J. B., Krakowsky J. M., Anderson K. P. A helix-loop-helix transcription factor-like gene is located at the mi locus. J Biol Chem. 1993 Oct 5;268(28):20687–20690. [PubMed] [Google Scholar]
  25. Imagawa M., Chiu R., Karin M. Transcription factor AP-2 mediates induction by two different signal-transduction pathways: protein kinase C and cAMP. Cell. 1987 Oct 23;51(2):251–260. doi: 10.1016/0092-8674(87)90152-8. [DOI] [PubMed] [Google Scholar]
  26. Jackson I. J., Chambers D. M., Tsukamoto K., Copeland N. G., Gilbert D. J., Jenkins N. A., Hearing V. A second tyrosinase-related protein, TRP-2, maps to and is mutated at the mouse slaty locus. EMBO J. 1992 Feb;11(2):527–535. doi: 10.1002/j.1460-2075.1992.tb05083.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kameyama K., Takemura T., Hamada Y., Sakai C., Kondoh S., Nishiyama S., Urabe K., Hearing V. J. Pigment production in murine melanoma cells is regulated by tyrosinase, tyrosinase-related protein 1 (TRP1), DOPAchrome tautomerase (TRP2), and a melanogenic inhibitor. J Invest Dermatol. 1993 Feb;100(2):126–131. doi: 10.1111/1523-1747.ep12462778. [DOI] [PubMed] [Google Scholar]
  28. Kikuchi H., Miura H., Yamamoto H., Takeuchi T., Dei T., Watanabe M. Characteristic sequences in the upstream region of the human tyrosinase gene. Biochim Biophys Acta. 1989 Dec 22;1009(3):283–286. doi: 10.1016/0167-4781(89)90115-2. [DOI] [PubMed] [Google Scholar]
  29. Klüppel M., Beermann F., Ruppert S., Schmid E., Hummler E., Schütz G. The mouse tyrosinase promoter is sufficient for expression in melanocytes and in the pigmented epithelium of the retina. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3777–3781. doi: 10.1073/pnas.88.9.3777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Kobayashi T., Urabe K., Winder A., Jiménez-Cervantes C., Imokawa G., Brewington T., Solano F., García-Borrón J. C., Hearing V. J. Tyrosinase related protein 1 (TRP1) functions as a DHICA oxidase in melanin biosynthesis. EMBO J. 1994 Dec 15;13(24):5818–5825. doi: 10.1002/j.1460-2075.1994.tb06925.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Kuzumaki T., Matsuda A., Wakamatsu K., Ito S., Ishikawa K. Eumelanin biosynthesis is regulated by coordinate expression of tyrosinase and tyrosinase-related protein-1 genes. Exp Cell Res. 1993 Jul;207(1):33–40. doi: 10.1006/excr.1993.1159. [DOI] [PubMed] [Google Scholar]
  32. Lowings P., Yavuzer U., Goding C. R. Positive and negative elements regulate a melanocyte-specific promoter. Mol Cell Biol. 1992 Aug;12(8):3653–3662. doi: 10.1128/mcb.12.8.3653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Ronai Z., Rutberg S., Yang Y. M. UV-responsive element (TGACAACA) from rat fibroblasts to human melanomas. Environ Mol Mutagen. 1994;23(3):157–163. doi: 10.1002/em.2850230302. [DOI] [PubMed] [Google Scholar]
  34. Weinberg R. A. The retinoblastoma protein and cell cycle control. Cell. 1995 May 5;81(3):323–330. doi: 10.1016/0092-8674(95)90385-2. [DOI] [PubMed] [Google Scholar]
  35. Wong G., Pawelek J. Melanocyte-stimulating hormone promotes activation of pre-existing tyrosinase molecules in Cloudman S91 melanoma cells. Nature. 1975 Jun 19;255(5510):644–646. doi: 10.1038/255644a0. [DOI] [PubMed] [Google Scholar]
  36. Yasumoto K., Mahalingam H., Suzuki H., Yoshizawa M., Yokoyama K. Transcriptional activation of the melanocyte-specific genes by the human homolog of the mouse Microphthalmia protein. J Biochem. 1995 Nov;118(5):874–881. doi: 10.1093/jb/118.5.874. [DOI] [PubMed] [Google Scholar]
  37. Yasumoto K., Yokoyama K., Shibata K., Tomita Y., Shibahara S. Microphthalmia-associated transcription factor as a regulator for melanocyte-specific transcription of the human tyrosinase gene. Mol Cell Biol. 1994 Dec;14(12):8058–8070. doi: 10.1128/mcb.14.12.8058. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Yavuzer U., Goding C. R. Melanocyte-specific gene expression: role of repression and identification of a melanocyte-specific factor, MSF. Mol Cell Biol. 1994 May;14(5):3494–3503. doi: 10.1128/mcb.14.5.3494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Yavuzer U., Keenan E., Lowings P., Vachtenheim J., Currie G., Goding C. R. The Microphthalmia gene product interacts with the retinoblastoma protein in vitro and is a target for deregulation of melanocyte-specific transcription. Oncogene. 1995 Jan 5;10(1):123–134. [PubMed] [Google Scholar]
  40. Yokoyama K., Suzuki H., Yasumoto K., Tomita Y., Shibahara S. Molecular cloning and functional analysis of a cDNA coding for human DOPAchrome tautomerase/tyrosinase-related protein-2. Biochim Biophys Acta. 1994 Apr 6;1217(3):317–321. doi: 10.1016/0167-4781(94)90292-5. [DOI] [PubMed] [Google Scholar]
  41. Yokoyama K., Yasumoto K., Suzuki H., Shibahara S. Cloning of the human DOPAchrome tautomerase/tyrosinase-related protein 2 gene and identification of two regulatory regions required for its pigment cell-specific expression. J Biol Chem. 1994 Oct 28;269(43):27080–27087. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES