Abstract
Genome-wide demethylation has been suggested to be a step in carcinogenesis1. Evidence for this notion comes from the frequently observed global DNA hypomethylation in tumour cells2, and from a recent study suggesting that defects in DNA methylation might contribute to the genomic instability of some colorectal tumour cell lines3. DNA hypomethylation has also been associated with abnormal chromosomal structures, as observed in cells from patients with ICF (Immunodeficiency, Centromeric instability and Facial abnormalities) syndrome4,5 and in cells treated with the demethylating agent 5-azadeoxycytidine6. Here we report that murine embryonic stem cells nullizygous for the major DNA methyltransferase (Dnmt1) gene exhibited significantly elevated mutation rates at both the endogenous hypoxanthine phosphoribosyltransferase (Hprt) gene and an integrated viral thymidine kinase (tk) transgene. Gene deletions were the predominant mutations at both loci. The major cause of the observed tk deletions was either mitotic recombination or chromosomal loss accompanied by duplication of the remaining chromosome. Our results imply an important role for mammalian DNA methylation in maintaining genome stability.
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References
Fearon, E. R. & Vogelstein, B. Agenetic model for colorectal tumorigenesis. Cell 61, 759–767 (1990).
Laird, P. W. & Jaenisch, R. The role of DNA methylation in cancer genetics and epigenetics. Annu. Rev. Genet. 30, 441–464 (1996).
Lengauer, C., Kinzler, K. W. & Vogelstein, B. DNA methylation and genetic instability in colorectal cancer cells. Proc. Natl Acad. Sci. USA 94, 2545–2550 (1997).
Jeanpierre, M. et al. An embryonic-like methylation pattern of classical satellite DNA is observed in ICF syndrome. Hum. Mol. Genet. 2, 731–735 (1993).
Ji, W. et al. DNA demethylation and pericentromeric rearrangements of chromosome 1. Mutat. Res. 379, 33–41 (1997).
Haaf, T. The effects of 5-azacytidine and 5-azadeoxycytidine on chromosome structure and function: implications for methylation-associated cellular processes. Pharmacol. Ther. 65, 19–46 (1995).
Li, E., Bestor, T. H. & Jaenisch, R. Targeted mutation of the DNA methyltransferase gene results in embryonic lethality. Cell 69, 915–926 (1992).
Lei, H. et al. De novo DNA cytosine methyltransferase activities in mouse embryonic stem cells. Development 122, 3195–3205 (1996).
Tucker, K. L. et al. Germ-line passage is required for establishment of methylation and expression patterns of imprinted but not of nonimprinted genes. Genes Dev. 10, 1008–1020 (1996).
Kendal, W. S. & Frost, P. Pitfalls and practice of Luria–Delbrück fluctuation analysis: a review. Cancer Res. 48, 1060–1065 (1988).
Shulman, M. J., Collins, C., Connor, A., Read, L. R. & Baker, M. D. Interchromosomal recombination is suppressed in mammalian somatic cells. EMBO J. 14, 4102–4107 (1995).
Jaenisch, R. DNA methylation and imprinting: why both? Trends Genet. 13, 323–329 (1997).
Petes, T. D., Malone, R. E. & Symington, L. S. in The Molecular and Cellular Biology of the Yeast Saccharomyces (eds Broach, J. R., Pringle, J. R. & Jones, E. W.) 407–521 (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, (1991)).
Proffitt, J. H., Davie, J. R., Swinton, D. & Hattman, S. 5-Methylcytosine is not detectable in Saccharomyces cerevisiae DNA. Mol. Cell. Biol. 4, 985–988 (1984).
Monk, M., Boubelik, M. & Lehnert, S. Temporal and regional changes in DNA methylation in the embryonic, extraembryonic and germ cell lineages during mouse embryo development. Development 99, 371–382 (1987).
Sanford, J. P., Clark, H. J., Chapman, V. M. & Rossant, J. Differences in DNA methylation during oogenesis and spermatogenesis and their persistence during early embryogenesis in the mouse. Genes Dev. 1, 1039–1046 (1987).
Maloisel, L. & Rossignol, J.-L. Suppression of crossing-over by DNA methylation in Ascobolus. Genes Dev. 12, 1381–1389 (1998).
Hsieh, C.-L. & Lieber, M. R. CpG methylated minichromosomes become inaccessible for V(D)J recombination after undergoing replication. EMBO J. 11, 315–325 (1992).
Engler, P., Weng, A. & Storb, U. Influence of CpG methylation and target splicing on V(D)J recombination in a transgenic substrate. Mol. Cell. Biol. 13, 571–577 (1993).
Yoder, J. A., Walsh, C. P. & Bestor, T. H. Cytosine methylation and the ecology of intragenomic parasites. Trends Genet. 13, 335–340 (1997).
Wahls, W. P., Wallace, L. J. & Moore, P. D. Hypervariable minisatellite DNA is a hotspot for homologous recombination in human cells. Cell 60, 95–103 (1990).
Shiroishi, T., Koide, T., Yoshino, M., Sagai, T. & Moriwaki, K. Hotspots of homologous recombination in mouse meiosis. Adv. Biophys. 31, 119–132 (1995).
Loeb, L. A. Mutator phenotype may be required for multistage carcinogenesis. Cancer Res. 51, 3075–3079 (1991).
Laird, P. W. et al. Suppression of intestinal neoplasia by DNA hypomethylation. Cell 81, 197–205 (1995).
Jackson-Grusby, L., Laird, P. W., Magge, S. N., Moeller, B. J. & Jaenisch, R. Mutagenicity of 5-aza-2′-deoxycytidine is mediated by the mammalian DNA methyltransferase. Porc. Natl. Acad. Sci. USA 94, 4681–4685 (1997).
Beard, C., Li, E. & Jaenisch, R. Loss of methylation activates Xist in somatic but not in embryonic cells. Genes Dev. 9, 2325–2334 (1995).
Melton, D. W., Konecki, D. S., Brennand, J. & Caskey, C. T. Structure, expression, and mutation of the hypoxanthine phosphoribosyltransferase gene. Proc. Natl. Acad. Sci. USA 81, 2147–2151 (1984).
Dietrich, W. F. et al. Agenetic map of the mouse with 4,006 simple sequence length polymorphisms. Nature Genet. 7, 220–245 (1994).
Acknowledgements
We thank P. P. Lee for pMT-3lox; W. M. Rideout III, and J. T. Lee for critical reading of the manuscript; and W. Thilly for helpful discussion. This work was supported by a grant from the swedish Medical Research Council (U.P.), postdoctoral fellowships from the Ann Fuller Fund (R.Z.C.) and the Cancer Research Fund of the Damon Runyon–Walter Winchell Foundation (L.J.-G.), and by the National Institutes of Health (R.Z.C., L.J.-G. and R.J.)
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Chen, R., Pettersson, U., Beard, C. et al. DNA hypomethylation leads to elevated mutation rates. Nature 395, 89–93 (1998). https://doi.org/10.1038/25779
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DOI: https://doi.org/10.1038/25779
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