Abstract
The MAGE1 gene codes for an antigen recognized on melanoma cell line MZ2-MEL by autologous cytolytic T lymphocytes. It is expressed in a number of tumors of different histological origins, but not in normal tissues except in testis. The MAGE1 promoter region was analyzed by performing transient transfections in MZ2-MEL cells with luciferase reporter plasmids. A fragment extending from nucleotide −792 to +118 exhibited high transcriptional activity. By deletional analysis of this fragment, we identified five activating regions designated C, A, B′, B, and D. The activity of region A depends on the presence of region B′ and vice versa. Two inverted Ets motifs contained in regions B′ and B were found to drive 90% of the activity of the MAGE1 promoter in MZ2-MEL cells. Electrophoretic mobility shift assays performed with a nuclear extract from MZ2-MEL cells and with competitor oligonucleotides containing an Ets consensus site showed that nuclear proteins bind to the Ets motif of regions B′ and B. Similar experiments suggested that region A binds transcription factors of the Sp1 family. The MAGE1 promoter was found to exert transcriptional activity in tumor cells where the MAGE1 gene is not expressed, suggesting that other mechanisms, such as demethylation, may contribute to the tumor-specific expression of the gene.
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Andrisani, O. M., Pot, D. A., Zu, Z., and Dixon, J. E. Three sequence-specific DNA-protein complexes are formed with the same promoter element essential for expression of the rat somatostatin gene. Mol Cell Biol 8: 1947–1956, 1988
Bassel-Duby, R., Hernandez, M. D., Gonzalez, M. A., Krueger, J. K., and Williams, R. S. A 40-kilodalton protein binds specifically to an upstream sequence element essential for muscle-specific transcription of the human myoglobin promoter. Mol Cell Biol 12: 5024–5032, 1992
Bird, A. The essentials of DNA methylation. Cell 70: 5–8, 1992
Boisclair, Y. R., Brown, A. L., Casola, S., and Rechler, M. M. Three clustered Sp1 sites are required for efficient transcription of the TATA-less promoter of the gene for insuline-like growth factor-binding protein-2 from the rat. J Biol Chem 268: 24892–24901, 1993
Brasseur, F., Marchand, M., Vanwijck, R., Hérin, M., Lethé, B., Chomez, P., and Boon, T. Human gene MAGE1, which codes for a tumor rejection antigen, is expressed by some breast tumors. Int J Cancer 52: 838–841, 1992
Brown, T. A. and McKnight, S. L. Specificities of protein-protein and protein-DNA interaction of GABPα and two newly defined ets-related proteins. Genes Dev 6: 2505–2512, 1992
Carthew, R. W., Chodosh, L. A., and Sharp, P. A. An RNA polymerase transcription factor binds to an upstream element in the adenovirus major late promoter. Cell 43: 439–448, 1985
De Plaen, E., Arden, K., Traversari, C., Gaforio, J. J., Szikora, J. P., De Smet, C., Brasseur, F., van der Bruggen, P., Lethé, B., Lurquin, C., Brasseur, R., Chomez, P., De Backer, O., Cavenee, W., and Boon, T. Structure, chromosomal localization, and expression of 12 genes of the MAGE family. Immunogenetics 40: 360–369, 1994
De Smet, C., Lurquin, C., van der Bruggen, P., De Plaen, E., Brasseur, F., and Boon, T. Sequence and expression pattern of the human MAGE2 gene. Immunogenetics 39: 121–129, 1994
De Wet, J. R., Wood, K. V., DeLuca, M., Helinski, D. R., and Subramani, S. Firefly luciferase gene: structure and expression in mammalian cells. Mol Cell Biol 7: 725–737, 1987
Dignam, J. D., Lebovitz, R. M., and Roeder, R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acid Res 11: 1475–1489, 1983
Eden, S. and Cedar, H. Role of DNA methylation in the regulation of transcription. Curr Opin Genet Dev 4: 255–259, 1994
Faisst, S. and Meyer, S. Compilation of vertebrate-encoded transcription factors. Nucleic Acid Res 20: 3–26, 1992
Gégonne, A., Bosselut, R., Bailly, R. A., and Ghysdael, J. Synergistic activation of the HTLV1 LTR Ets-responsive region by transcription factors Ets1 and Sp1. EMBO J 12: 1169–1178, 1993
Gunther, C. V., Nye,J. A., Bryner, R. S., and Graves, B. J. Sequence-specific DNA binding of the proto-oncoprotein ets-1 defines a transcriptional activator sequence within the long terminal repeat of the Moloney murine sarcoma virus. Genes Dev 4: 667–679, 1990
Hérin, M., Lemoine, C., Weynants, P., Vessière, F., Van Pel, A., Knuth, A., Devos, R., and Boon, T. Production of stable cytolytic T-cell clones directed against autologous human melanoma. Int J Cancer 39: 390–396, 1987
Jones, P. A. and Buckley, J. D. The role of DNA methylation in cancer. Adv Cancer Res 54: 1–23, 1990
Lemaigre, F. P., Courtois, S. J., Lafontaine, D. A., and Rousseau, G. G. Evidence that the upstream stimulatory factor and the Sp1 transcription factor bind in vitro to the promoter of the human-growth hormone gene. Eur J Biochem 181: 555–561, 1989
MacGregor, G. R., Mogg, A. E., Burke, J. F., and Caskey, C. T. Histochemical staining of clonal mammalian cell lines expressing E. Coli β-galactosidase indicates heterogeneous expression of the bacterial gene. Somat Mol Cell Genet 13: 253–265, 1987
Muscatelli, F., Walker, A. P., De Plaen, E., Stafford, A. N., and Monaco, A. P. Isolation and characterization of a new MAGE gene family in the Xp21.3 region. Proc Natl Acad Sci USA 9295: 4987–4991, 1995
Nordeen, S. K. Luciferase reporter gene vectors for analysis of promoters and enhancers. BioTechniques 6: 454–457, 1988
Patard, J.-J., Brasseur, F., Gil-Diez, S., Radvany, F., Marchand, M., François, P., Abi-Aad, A., Van Cangh, P., Abbou, C. C., Chopin, D., and Boon, T. Expression of MAGE genes in transitional-cell carcinomas of the urinary bladder. Int J Cancer 64: 60–64, 1995
Philippe, M., Larondelle, Y., Lemaigre, F., Mariamé, B., Delhez, H., Mason, P., Luzzato, L., and Rouseau, G. G. Promoter function of the human glucose-6-phosphate dehydrogenase gene depends on two GC boxes that are cell specifically controlled. Eur J Biochem 226: 377–384, 1994
Postel, E. H., Mango, S. E., and Flint, S. J. A nuclease-hypersensitive element of the human c-myc promoter interacts with a transcription initiation factor. Mol Cell Biol 9: 5123–5133, 1989
Rosenthal, N. Identification of regulatory elements of cloned genes with functional assays. Methods Enzymol 152: 704–720, 1987
Sambrook, J., Fritsch, E. F., and Maniatis, T. Molecular Cloning: A Laboratory Manual, 2nd edn, Cold Spring Harbor Laboratory, Cold Spring Harbor, 1989
Traversari, C., van der Bruggen, P., Van den Eynde, B., Hainaut, P., Lemoine, C., Ohta, N., Old, L., and Boon, T. Transfection and expression of a gene coding for a human melanoma antigen recognized by autologous cytolytic T lymphocytes. Immunogenetics 35: 145–152, 1992
Ven den Eynde, B., Hainaut, P., Hérin, M., Knuth, A., Lemoine, C., Weynants, P., van der Bruggen, P., and Boon, T. Presence on a human melanoma of multiple antigens recognized by autologous CTL. Int J Cancer 44: 634–640, 1989
Van den Eynde, B., Peeters, O., De Backer, O., Gaugler, B., Lucas, S., and Boon, T. A new family of genes coding for an antigen recognized by autologous cytolytic T lymphocytes on a human melanoma. J Exp Med in press, 1995
Van der Bruggen, P., Traversari, C., Chomez, P., Lurquin, C., De Plaen, E., Van den Eynde, B., Knuth, A., and Boon, T. A gene encoding an antigen recognized by cytolytic T lymphocytes on a human melanoma. Science 254: 1643–1647, 1991
Wasylyk, B., Hahn, L., and Giovanne, A. The Ets family of transcription factors. Eur J Biochem 211: 7–18, 1993
Weber, J., Salgaller, M., Samid, D., Johnson, B., Herlyn, M., Lassam, N., Treisman, J., and Rosenberg, S. A. Expression of the MAGE1 tumor antigen is upregulated by the demethylating agent 5-aza-2′-deoxycytidine. Cancer Res 54: 1766–1771, 1994
Weynants, P., Lethé, B., Brasseur, F., Marchand, M., and Boon, T. Expression of MAGE genes by non-small-cell lung carcinomas. Int J Cancer 56: 826–829, 1994
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De Smet, C., Courtois, S.J., Faraoni, I. et al. Involvement of two Ets binding sites in the transcriptional activation of the MAGE1 gene. Immunogenetics 42, 282–290 (1995). https://doi.org/10.1007/BF00176446
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DOI: https://doi.org/10.1007/BF00176446