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O6-Methylguanine repair of methylated DNA in vitro: Cell cycle-dependence of rat liver methyltransferase activity

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  • Experimental Oncology
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Summary

O6-Methylguanine DNA transferase activity was investigated in liver proteins obtained at various intervals after partial hepatectomy and/or after hydroxyurea-induced synchronization of the liver cell cycle. Liver proteins were incubated with 3H-methylated calf thymus DNA as previously described by Pegg et al. (1981). The loss of O6-methylguanine was measured by radiochromatography of DNA hydrolysates. The extent of O6-methylguanine repair differed during the cell cycle: the activity increased in late G1, reached a maximum in early S phase and declined in late S phase and G2M. These results indicate that hepatocytes are endowed with an increased DNA repair capacity for this promutagenic lesion during the period of highest transformation sensitivity in the cell cycle. Though increased, however, this repair potential does not, because of its exhaustibility, appear to be sufficient to prevent initiation of transformation after high doses of alkylating carcinogens.

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References

  • Abbott PJ, Saffhill R (1979) DNA synthesis with methylated poly(dC-dG) templates: evidence for a competitive nature to miscoding by O6-methylguanine. Biochim Biophys Acta 562:51–61

    Google Scholar 

  • Andrae U, Greim H (1979) Induction of DNA repair replication by hydroxyurea in human lymphoblastoid cells mediated by liver microsomes and NADPH. Biochem Biophys Res Commun 87:50–58

    Google Scholar 

  • Bertram JS, Heidelberger C (1974) Cell cycle dependency of oncogenic transformation induced by N-methyl-N′-nitro-N-nitrosoguanidine in culture. Cancer Res 34:526–537

    Google Scholar 

  • Boehm TLJ, Drahovsky D (1981) Hypomethylation of DNA in Raji cells after treatment with N-methyl-N-nitrosourea. Carcinogenesis 2:39–42

    Google Scholar 

  • Bucher NLR, Malt RA (1971) Regeneration of liver and kidney. Little Brown, Boston

    Google Scholar 

  • Burton K (1956) A study of the conditions and mechanisms of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J 62:315–323

    Google Scholar 

  • Chu YH, Craig AW, O'Connor PJ (1981) Repair of O6-methylguanine in rat liver DNA is enhanced by pretreatment with single or multiple doses of aflatoxin B1. Br J Cancer 43:850–855

    Google Scholar 

  • Cooper GM (1982) Cellular transforming genes. Science 217:801–806

    Google Scholar 

  • Cooper DP, O'Connor PJ, Margison GP (1982) Effect of acute doses of 2-acetylaminofluorene on the capacity of rat liver to repair methylated purines in DNA in vivo and in vitro. Cancer Res 42:4203–4209

    Google Scholar 

  • Gombar CT, Katz EJ, Magee PN, Sirover MA (1981) Induction of the DNA repair enzymes uracil DNA glycosylase and 3-methyladenine DNA glycosylase in regenerating rat liver. Carcinogenesis 2:595–599

    Google Scholar 

  • Goth R, Rajewsky MF (1974) Molecular and cellular mechanisms associated with pulse carcinogenesis in the rat nervous system by ethylnitrosourea: ethylation of nucleic acids and elimination rates of ethylated bases from the DNA of different tissues. Z Krebsforsch 82:37–64

    Google Scholar 

  • Gupta PK, Sirover MA (1981) Cell cycle regulation of DNA repair in normal and repair deficient human cells. Chem Biol Interact 36:19–31

    Google Scholar 

  • Higgins GM, Anderson RM (1931) Experimental pathology of the liver of the white rat following partial surgical removal. Arch Pathol 12:186–202

    Google Scholar 

  • Kleinhues P, Margison GP (1974) Carcinogenicity of N-methyl-N-nitrosourea: Possible role of excision repair of O6-methylguanine from DNA. J Natl Cancer Inst 53:1839–1841

    Google Scholar 

  • Loveless A (1969) Possible relevance of O6-alkylation of deoxyguanosine to the mutagenicity and carcinogenicity of nitrosamines and nitrosamides. Nature 223:206–207

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin Phenol reagent. J Biol Chem 193:265–275

    Google Scholar 

  • Margison GP (1981) Effect of pretreatment with N-methyl-N-nitrosourea on the repair of O6-methylguanine in liver DNA. Carcinogenesis 2:431–434

    Google Scholar 

  • Montesano R, Brésil H, Planche-Martel G, Margison GP, Pegg AE (1980) Effect of chronic treatment of rats with dimethylnitrosamine on the removal of O6-methylguanine from DNA. Cancer Res 40:452–458

    Google Scholar 

  • Müller L, Rabes HM (1983) Cell cycle-dependent initiation of ATP-ase-deficient preneoplastic cell populations in regenerating rat liver. J Cancer Res Clin Oncol 105:A 22

    Google Scholar 

  • Nicoll JW, Swann PF, Pegg AE (1975) Effect of dimethylnitrosamine on persistence of methylated guanines in rat liver and kidney DNA. Nature 254:261–262

    Google Scholar 

  • Pegg AE, Perry W (1981) Stimulation of transfer of methyl groups from O6-methylguanine in DNA to protein by rat liver extracts in response to hepatotoxins. Carcinogenesis 11:1195–1200

    Google Scholar 

  • Pegg AE, Wiest L (1983) Regulation of O6-methylguanine-DNA methyltransferase levels in rat liver and kidney. Cancer Res 43:972–975

    Google Scholar 

  • Pegg AE, Perry W, Bennett RA (1981) Effect of partial hepatectomy on removal of O6-methylguanine from alkylated DNA by rat liver extracts. Biochem J 197:195–201

    Google Scholar 

  • Pegg AE, Roberfroid M, v. Bahr C, Foote RS, Mitra S, Brésil H, Likhachev A, Montesano R (1982) Removal of O6-methylguanine from DNA by human liver fractions. Proc Natl Acad Sci USA 79:5162–5165

    Google Scholar 

  • Pegg AE, Wiest L, Foote RS, Mitra S, Wayne P (1983) Purification and properties of O6-methylguanine-DNA transmethylase from rat liver. J Biol Chem 258:2327–2333

    Google Scholar 

  • rabes HM (1979) Proliferative Vorgänge während der Frühstadien der malignen Transformation. Verh Dtsch Ges Pathol 63:18–39

    Google Scholar 

  • Rabes HM, Tuczek HV (1970) Quantitative autoradiographische Untersuchung zur Heterogenität der Leberzellproliferation nach partieller Hepatektomie. Virchows Arch Abt B Zellpathol 6:302–312

    Google Scholar 

  • Rabes HM, Iseler G, Czichos S, Tuczek HV (1977) Synchronization of hepatocellular DNA synthesis in regenerating rat liver by continuous infusion of hydroxyurea. Cancer Res 37:1105–1111

    Google Scholar 

  • Rabes HM, Kerler R, Wilhelm R, Rode G, Riess H (1979) Alkylation of DNA and RNA by (14C)dimethylnitrosamine injhydroxyurea-synchronized regenerating rat liver. Cancer Res 39:4228–4236

    Google Scholar 

  • Rabes HM, Wilhelm R, Kerler R, Rode G (1982) Dose- and cell cycle-dependent O6-methylguanine elimination from DNA in regenerating rat liver after (14C)dimethylnitrosamine injection. Cancer Res 42:3814–3821

    Google Scholar 

  • Rabes HM, Kerler R, Wilhelm R (1983) DNA alkylation and tumor induction in regenerating rat liver after cell cycle-related continuous N-nitrosodimethylamine infusion. J Natl Cancer Inst 70:193–198

    Google Scholar 

  • Rabes HM, Kerler R, Rode G, Schuster C, Wilhelm R (1984) O6-methylguanine repair in liver cells in vivo: Comparison between G1 and S-phase of the cell cycle. J Cancer Res Clin Oncol 108:36–45

    Google Scholar 

  • Rabes HM, Kerler R, Schuster C, Rode G, Legner M, Müller L, Bücher T (1985) Cell proliferation and DNA repair in the liver during early stages of chemical carcinogenesis. In: Weber G (ed) Advances in enzyme regulation vol 23. Pergamon Press, Oxford, New York, pp 241–262

    Google Scholar 

  • Riggs AD, Jones PA (1983) 5-Methylcytosine, gene regulation and cancer. Adv Cancer Res 40:1–30

    Google Scholar 

  • Sirover MA (1979) Induction of the DNA repair enzyme uracil DNA glycosylase in stimulated human lymphocytes. Cancer Res 39:2090–2095

    Google Scholar 

  • Sukumar S, Notario V, Martin-Zanca D, Barbacid M (1983) Induction of mammary carcinomas in rats by nitrosomethyl-urea involves the malignant activation of the H-ras-1 locus by single point mutations. Nature 306:658–662

    Google Scholar 

  • Swann PF, Magee PN (1968) Nitrosamine-induced carcinogenesis. The alkylation of nucleic acids of the rat by N-methyl-N-nitrosourea, dimethylnitrosamine, dimethyl sulphate and methyl methanesulphonate. Biochem J 110:39–47

    Google Scholar 

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Supported by grants from Deutsche Forschungsgemeinschaft

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Schuster, C., Rode, G. & Rabes, H.M. O6-Methylguanine repair of methylated DNA in vitro: Cell cycle-dependence of rat liver methyltransferase activity. J Cancer Res Clin Oncol 110, 98–102 (1985). https://doi.org/10.1007/BF00402719

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  • DOI: https://doi.org/10.1007/BF00402719

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