Abstract
A cloned DNA fragment (λR31) containing the human gene for melanoma‐associated ME491 antigen was transfected into mouse fibroblast cell lines and the antigen expression was studied. Our preliminary observation of higher expression of the antigen in more malignant Ltk cells and weaker expression in less malignant NIH3T3 cells tempted us to investigate the antigen expression in Harvey (H)‐ras‐transformed NIH3T3 cells. It was observed that malignant transformation of the λR31‐transfected NIH3T3 cells by H‐ras oncogene enhanced the antigen expression to some extent. Northern blot analysis suggested that the enhancement occurred at the transcriptionsl level. Nucleotide sequence analysis of the 5′‐regulatory region of the ME491 antigen gene in λR31 identified a number of consensus sequence motifs for binding of transcription factors such as Sp1, AP‐2 and polyomavirus enhancer binding proteins 2 and 3. A consensus sequence motif for binding of AP‐1, known as a ras‐responsive element, was not found in that region. The significance and possible involvement of the transcription factors in the enhancement of ME491 antigen expression are discussed.
Keywords: Melanoma‐associated antigen, Human gene, Gene regulation, Malignant transformation
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REFERENCES
- 1. ) Clark , W. H. , Jr. , Elder , D. E. , Guerry , D.IV , Epstein , M. N. , Greene , M. H. and van Horn , M.A study of tumor progression: the precursor lesions of superficial spreading and nodular melanoma . Human Pathol. , 15 , 1147 – 1165 ( 1984. ). [DOI] [PubMed] [Google Scholar]
- 2. ) Elder , D. E. and Clark , W. H. , Jr.Developmental biology of malignant melanoma . In “ Pigment Cell, Vol. 8 ,” ed Elder D. E. , pp. 1 – 28 ( 1987. ). Karger; , Basel ). [Google Scholar]
- 3. ) Elder , D. E. and Clark , W. H. , Jr.Tumor progression and prognosis in malignant melanoma . In “ Pigment Cell, Vol. 8 ,” ed Elder D. E. , pp. 51 – 80 ( 1987. ). Karger; , Basel ). [Google Scholar]
- 4. ) Herlyn , M. , Clark , W. H. , Rodeck , U. , Mancianti , M. L. , Jambrosic , J. and Koprowski , H.Biology of tumor progression in human melanocytes . Lab. Invest. , 56 , 461 – 474 ( 1987. ). [PubMed] [Google Scholar]
- 5. ) Herlyn , M. and Koprowski , H.Melanoma antigens: immunological and biological characterization and clinical significance . Ann. Rev. Immunol. , 6 , 283 – 308 ( 1988. ). [DOI] [PubMed] [Google Scholar]
- 6. ) Holzmann , B. , Brocker , E. B. , Lehmann , J. M. , Ruiter , D. J , , Sorg , C. , Riethmuller , G. and Johnson , J. P.Tumor progression in human malignant melanoma: five stages defined by their antigenic phenotypes . Int. J. Cancer , 39 , 466 – 471 ( 1987. ). [DOI] [PubMed] [Google Scholar]
- 7. ) Atkinson , B. , Ernst , C. S. , Ghrist , B. F. D. , Ross , A. H. , Clark , W. H , Herlyn , M , Herlyn , D.Maul , G. , Steplewski , Z. and Koprowski , H.Monoclonal antibody to a highly glycosylated protein reacts in fixed tissue with melanoma and other tumors . Hybridoma , 4 , 243 – 255 ( 1985. ). [DOI] [PubMed] [Google Scholar]
- 8. ) Hotta , H. , Ross , A. H , Huebner , K. , Isobe , M , Wendeborn , S. , Chao , M. V. , Ricciardi , R. P. , Tsujimoto , Y. , Croce , C. M. and Koprowski , H.Molecular cloning and characterization of an antigen associated with early stages of melanoma tumor progression . Cancer Res. , 48 , 2955 – 2962 ( 1988. ). [PubMed] [Google Scholar]
- 9. ) Colbere‐Garapin , F. , Chousterman , S. , Horodniceanu , F. , Kourilski , P. and Qarapin , A. C.Cloning of the active thymidine kinase gene of herpes simplex virus type 1 in Escherichia coli K‐12 . Proc. Natl Acad. Sci. USA , 76 , 3755 – 3759 ( 1979. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 10. ) Southern , P. J. and Berg , P.Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter . J. Mol. Appl. Genet. , 1 , 327 – 341 ( 1982. ). [PubMed] [Google Scholar]
- 11. ) Santos , E. , Tronick , S. R. , Aaronson , S. A. , Pulciani , S. and Barbacid , M.T24 human bladder carcinoma oncogene is an activated form of the normal human homologue of BALB‐ and Harvey‐MSV transforming genes . Nature , 298 , 343 – 347 ( 1982. ). [DOI] [PubMed] [Google Scholar]
- 12. ) Perucho , M. , Goldfarb , M. , Shimizu , K. , Lama , C. , Fogh , J. and Wigler , M.Human‐tumor‐derived cell lines contain common and different transforming genes . Cell , 27 , 467 – 476 ( 1981. ). [DOI] [PubMed] [Google Scholar]
- 13. ) Blair , D. G. , Cooper , C. S. , Oskarsson , M. K. , Eader , L. A. and Vande Woude , G. F.New method for detecting cellular transforming genes . Science , 218 , 1122 – 1125 ( 1982. ). [DOI] [PubMed] [Google Scholar]
- 14. ) Bernstein , S. C. and Weinberg , R. A.Expression of the metastatic phenotype in cells transfected with human metastatic tumor DNA . Proc. Natl. Acad. Sci. USA , 82 , 1726 – 1730 ( 1985. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 15. ) Wigler , M. , Pellicer , A. , Silverstein , S. , Axel , R. , Urlaub , G. and Chasin , L.DNA‐mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells . Proc. Natl. Acad. Sci. USA , 76 , 1373 – 1376 ( 1979. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 16. ) Chao , M. V. , Bothwell , M. A. , Ross , A. H. , Koprowski , H. , Lanahan , A. A. , Buck , C. R. and Sehgal , A.Gene transfer and molecular cloning of the human NGF receptor . Science , 232 , 518 – 521 ( 1986. ). [DOI] [PubMed] [Google Scholar]
- 17. ) Goding , J. W.The chromic chloride method of coupling antigens to erythrocytes: definition of some important parameters . J. Immunol. Methods , 10 , 61 – 66 ( 1976. ). [DOI] [PubMed] [Google Scholar]
- 18. ) Ar‐Rushdi , A. , Tan , K. B. and Croce , C. M.Tran‐scriptional control of the expression of mouse globin genes in myeloma × erythroleukemia cell hybrids . Sotnat. Cell Genet. , 8 , 151 – 161 ( 1982. ). [DOI] [PubMed] [Google Scholar]
- 19. ) Davis L. G. , Dibner M. D. and Battey J. F. ( ed. ) ) “ Basic Methods in Molecular Biology ” ( 1986. ). Elsevier Science Publishing Co., Inc. , New York . [Google Scholar]
- 20. ) Maniatis T. , Fritsch E. F. and Sambrook J. ( ed. ) “ Molecular Cloning: A Laboratory Manual ” ( 1982. ). Cold Spring Harbor Laboratory; , Cold Spring Harbor ). [Google Scholar]
- 21. ) Lemischka , I. R. , Farmer , S. , Racaniello , V. R. and Sharp , P. A.Nucleotide sequence and evolution of a mammalian a‐tubulin messenger RNA . J. Mol. Biol. , 151 , 101 – 120 ( 1981. ). [DOI] [PubMed] [Google Scholar]
- 22. ) Sanger , F. , Nicklen , S. and Coulson , A. R.DNA sequencing with chain‐terminating inhibitors . Proc. Natl. Acad. Sci. USA , 74 , 5463 – 5467 ( 1977. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 23. ) Briggs , M. R. , Kadonaga , J. T. , Bell , S. P. and Tjian , R.Purification and biochemical characterization of the promoter‐specific transcription factor, Spl . Science , 234 , 47 – 52 ( 1986. ). [DOI] [PubMed] [Google Scholar]
- 24. ) Ishii , S. , Merlino , G. T. and Pastan , LPromoter region of the human Harvey ras proto‐oncogene: similarity to the EGF receptor proto‐oncogene promoter . Science , 230 , 1378 – 1381 ( 1985. ). [DOI] [PubMed] [Google Scholar]
- 25. ) Ishii , S. , Xu , Y‐H. , Stratton , R. H. , Roe , B. A. , Merlino , G. T. and Pastan , LCharacterization and sequence of the promoter region of the human epidermal growth factor receptor gene . Proc. Natl. Acad. Sci, USA , 82 , 4920 – 4924 ( 1985. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 26. ) Imagawa , M. , Chiu , R. and Karin , M.Transcription factor AP‐2 mediates induction by two different signal‐transduction pathways: protein kinase C and cAMP . Cell , 51 , 251 – 260 ( 1987. ). [DOI] [PubMed] [Google Scholar]
- 27. ) Satake , M. , Ibaraki , T. and Ito , Y.Modulation of polyomavirus enhancer binding proteins by Ha‐ras oncogene . Oncogene , 3 , 69 – 78 ( 1988. ). [Google Scholar]
- 28. ) Yamaguchi , Y. , Satake , M. and Ito , Y.Two overlapping sequence motifs within the polyomavirus enhancer are independently the targets of stimulation by both the tumor promoter 12‐O‐tetradecanoylphorbol‐13‐acetate and the Ha‐ras oncogene . J. Viral. , 63 , 1040 – 1048 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 29. ) Wasylyk , C. , Imler , J. L. , Perez‐Mutul , J. and Wasylyk , B.The c‐Ha‐ras oncogene and a tumor promoter activate the polyoma virus enhancer . Cell , 48 , 525 – 534 ( 1987. ). [DOI] [PubMed] [Google Scholar]
- 30. ) Imler , J. L. , Schatz , C. , Wasylyk , C. , Chatton , B. and Wasylyk , B.A Harvey‐ras responsive transcription element is also responsive to a tumour‐promoter and to serum . Nature , 332 , 275 – 278 ( 1988. ). [DOI] [PubMed] [Google Scholar]
- 31. ) Schönthal , A. , Herrlich , P. , Rahmsdorf , H. J. and Ponta , H.Requirement for fos gene expression in the transcriptional activation of collagenase by other oncogenes and phorbol esters . Cell , 54 , 325 – 334 ( 1988. ). [DOI] [PubMed] [Google Scholar]
- 32. ) Flier , J. S. , Mueckler , M. M. , Usher , P. and Lodish , H. F.Elevated levels of glucose transport and transporter messenger RNA are induced by ras or src oncogenes . Science , 235 , 1492 – 1498 ( 1987. ). [DOI] [PubMed] [Google Scholar]
- 33. ) Hume , C. R. , Accolla , R. S. and Lee , J. S.Defective HLA class II expression in a regulatory mutant is partially complemented by activated ras oncogenes . Proc. Natl Acad. Sci USA , 84 , 8603 – 8607 ( 1987. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 34. ) Owen , R. D. and Ostrowski , M. C.Rapid and selective alterations in the expression of cellular genes accompany conditional transcription of Ha‐v‐ras in NIH3T3 cells . Mol. Cell. Biol. , 7 , 2512 – 2520 ( 1987. ). [DOI] [PMC free article] [PubMed] [Google Scholar]