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Silencing of metallothionein-I gene in mouse lymphosarcoma cells by methylation

Oncogene volume 18, pages 6287–6295 (1999)Cite this article

Abstract

Metallothionein-I (MT-I) gene is silenced by methylation of CpG islands in mouse lymphosarcoma P1798 cells but not in the thymus, the cell type from which the tumor was derived. Bisulfite genomic sequencing revealed that all 21 CpG dinucleotides present within −216 bp to +1 bp with respect to transcription start site are methylated in the tumor cell line, but none is methylated in the thymus. The lymphosarcoma cells induced MT-I in response to heavy metals only after demethylation with 5-azacytidine (5-AsaC). The electrophoretic mobility shift assay using specific oligonucleotide probes showed that the key transcription factors regulating MT-I gene (e.g., MTF-1, Sp 1 and MLTF/USF) are active in P1798 cells. In vivo footprinting of the proximal promoter region showed that none of the metal regulatory elements (MREs) or MLTF/USF are occupied in response to heavy metals. Demethylation of the lymphosarcoma cells with 5-AzaC resulted in constitutive footprinting at MLTF/ARE, and zinc-inducible footprinting at MRE-c, MRE-d and MRE-e sites. Demethylation of just 10 – 20% of the CpG islands was sufficient to render the gene inducible by cadmium or zinc. The MT-I induction persisted in the cancer cells for several generations even after withdrawal of 5-AzaC from the culture medium.

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References

  • Aniskovitch LP and Jacob ST. . 1997 Arch. Biochem. Biophys. 341: 337–346.

  • Aniskovitch LP and Jacob ST. . 1998 Oncogene 16: 1475–1486.

  • Aschner M, Cherian MG, Klaassen CD, Palmiter RD, Erickson JC and Bush AI. . 1997 Toxicol. Appl. Pharmacol. 142: 229–242.

  • Baylin SB. . 1997 Science 277: 1948–1949.

  • Baylin SB, Herman JG, Graff JR, Vertino PM and Issa JP. . 1998 Adv. Cancer Res. 72: 141–196.

  • Cameron EE, Bachman KE, Myohanen S, Herman JG and Baylin SB. . 1999 Nat. Genet. 21: 103–107.

  • Carthew RW, Chodosh LA and Sharp PA. . 1987 Genes Dev. 1: 973–980.

  • Chomczynski P and Sacchi N. . 1987 Anal. Biochem. 162: 156–159.

  • Chubatsu LS and Meneghini R. . 1993 Biochem. J. 291: 193–198.

  • Clark SJ, Harrison J and Frommer M. . 1995 Nat. Genet. 10: 20–27.

  • Clark SJ, Harrison J, Paul CL and Frommer M. . 1994 Nucleic Acids Res. 22: 2990–2997.

  • Compere SJ and Palmiter RD. . 1981 Cell 25: 233–240.

  • Datta PK and Jacob ST. . 1993 Cell. Mol. Biol. Res. 39: 439–449.

  • Datta PK and Jacob ST. . 1997 Biochem. Biophys. Res. Commun. 230: 159–163.

  • Deng J and Szyf M. . 1998 J. Biol. Chem. 273: 22869–22872.

  • Feinberg AP, Gehrke CW, Kuo KC and Ehrlich M. . 1988 Cancer Res. 48: 1159–1161.

  • Frommer M, McDonald LE, Millar DS, Collis CM, Watt F, Grigg GW, Molloy PL and Paul CL. . 1992 Proc. Natl. Acad. Sci. USA 89: 1827–1831.

  • Ghoshal K and Jacob ST. . 1996 Biochem J. 317: 689–695.

  • Ghoshal K, Li Z and Jacob ST. . 1998 Proc. Natl. Acad. Sci. USA 95: 10390–10395.

  • Ghoshal K, Wang Y, Sheridan J and Jacob ST. . 1998 J. Biol. Chem. 277: 27904–27910.

  • Harrison J, Stirzaker C and Clark SJ. . 1998 Anal. Biochem. 264: 129–132.

  • Heuchel R, Radtke F, Georgiev O, Stark G, Aguet M and Schaffner W. . 1994 EMBO J. 13: 2870–2875.

  • Jaenisch R. . 1997 Trends Genet. 13: 323–329.

  • Kagi JA. . 1991 Methods Enzymol. 205: 613–626.

  • Kass SU, Pruss D and Wolffe AP. . 1997 Trends Genet. 13: 444–449.

  • Kelley SL, Basu A, Teicher BA, Hacker MP, Hamer DH and Lazo JS. . 1988 Science 241: 1813–1815.

  • Klein CB and Costa M. . 1997 Mutat. Res. 386: 163–180.

  • Knight SJ, Flannery AV, Hirst MC, Campbell L, Christodoulou Z, Phelps SR, Pointon J, Middleton-Price HR, Barnicoat A, Pembrey ME, et al. 1993 Cell 74: 127–134.

  • Lengauer C, Kinzler KW and Vogelstein B. . 1997 Proc. Natl. Acad. Sci. USA 94: 2545–2550.

  • Lewin B. . 1998 Cell 93: 301–303.

  • Lie E, Beard C, Forster AC, Bestor TH and Jaenisch R. . 1993 Cold Spring Harb. Symp. Quant. Biol. 58: 297–305.

  • Li Q, Hu N, Daggett MA, Chu WA, Bittel D, Johnson JA and Andrews GK. . 1998 Nucleic Acids Res. 26: 5182–5189.

  • Mueller PR and Wold B. . 1989 Science 246: 780–786.

  • Nan X, Campoy FJ and Bird A. . 1997 Cell 88: 471–481.

  • Ng MH, Chung YF, Lo KW, Wickham NW, Lee JC and Huang DP. . 1997 Blood 89: 2500–2506.

  • Norris DP, Patel D, Kay GF, Penny GD, Brockdorff N, Sheardown SA and Rastan S. . 1994 Cell 77: 41–51.

  • Palmiter RD. . 1987 EXS 52: 63–80.

  • Ping D, Jones PL and Boss JM. . 1996 Immunity 4: 455–469.

  • Pitt BR, Schwarz M, Woo ES, Yee E, Wasserloos K, Tran S, Weng W, Mannix RJ, Watkins SA, Tyurina YY, Tyurin VA, Kagan VE and Lazo JS. . 1997 Am. J. Physiol. 273: L856–L865.

  • Quaife CJ, Findley SD, Erickson JC, Froelic GJ, Kelly EJ, Zambrowicz BP and Palmiter RD. . 1994 Biochemistry 33: 7250–7259.

  • Radtke F, Heuchel R, Georgiev O, Hergersberg M, Gariglio M, Dembic Z and Schaffner W. . 1993 EMBO J. 12: 1355–1362.

  • Radtke F, Hug M, Georgiev O, Matsuo K and Schaffner W. . 1996 Biol. Chem. Hoppe Seyler 377: 47–56.

  • Robertson KD, Hayward SD, Ling PD, Samid D and Ambinder RF. . 1995 Mol. Cell. Biol. 15: 6150–6159.

  • Santi DV, Garrett CE and Barr PJ. . 1983 Cell 33: 9–10.

  • Santi DV, Norment A and Garrett CE. . 1984 Proc. Natl. Acad. Sci. USA 81: 6993–6997.

  • Schwartz MA, Lazo JS, Yalowich JC, Allen WP, Whitmore M, Bergonia HA, Tzeng E, Billiar TR, Robbins PD, Lancaster Jr JR et al. 1995 Proc. Natl. Acad. Sci. USA 92: 4452–4456.

  • Szyf M. . 1996 Pharmacol. Ther. 70: 1–37.

  • Warnecke PM, Stirzacker C, Melki JR, Millar DS, Paul CL and Clark SJ. . 1997 Nucleic Acids Res. 25: 4422–4426.

  • Watt F and Molloy PL. . 1990 Philos. Trans. R. Soc. Lond. B. Biol. Sci. 326: 267–275.

  • Wood KM and Thompson Jr AE. . 1984 Mol. Cell. Endocrinol. 37: 169–180.

  • Yagle MK and Palmiter RD. . 1985 Mol. Cell. Biol. 5: 291–294.

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Acknowledgements

The authors are grateful to Aubrey Thompson and Richard Palmiter for providing mouse lymphosarcoma P1798 cells and mouse MT-I minigene, respectively. This work was supported, in part, by the USPHS Grant CA61321 to ST Jacob and ACS Institutional Research Grant to K Ghoshal.

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Authors and Affiliations

  1. Department of Medical Biochemistry, College of Medicine, 333 Hamilton Hall, 1645 Neil Avenue, The Ohio State University, Columbus, OH 43210, Ohio, USA

    Sarmila Majumder, Kalpana Ghoshal, Zhiling Li & Samson T Jacob

  2. Comprehensive Cancer Center, The Ohio State University, Columbus, 43210, Ohio, OH, USA

    Yuan Bo

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  1. Sarmila Majumder
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  2. Kalpana Ghoshal
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  4. Yuan Bo
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  5. Samson T Jacob
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Majumder, S., Ghoshal, K., Li, Z. et al. Silencing of metallothionein-I gene in mouse lymphosarcoma cells by methylation. Oncogene 18, 6287–6295 (1999). https://doi.org/10.1038/sj.onc.1203004

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