Summary
Comparisons of nucleic acid methylation between paired neoplastic and non-neoplastic mouse cell lines have shown a striking difference in the deoxyribonucleic acid (DNA) peak eluted from methylated albumin-kieselguhr columns (R. Gantt and V. J. Evans, 1969, Cancer Res. 29: 536–541). Since mouse satellite DNA is relatively highly methylated, its 5-methylcytosine content was compared with mainband DNA in these two paired cell lines to determine whether this might account for the observed differences. The cell DNA was labeled with methyl-labeled methionine and isolated from the cells by repeated neutral cesium chloride isopycnic centrifugation. The satellite DNA strands were then separated in an alkaline cesium chloride gradient. Both the 5-methylcytosine content and the relative amounts of satellite DNA were indistinguishable in the paired cell lines. Further, the results showed that both strands of satellite DNA had virtually equal amounts of 5-methylcytosine, although the heavy strand contains 1.5 times more cytosine than the light strand.
Similar content being viewed by others
References
Gold, M., J. Hurwitz, and M. Anders. 1963. The enzymatic methylation of RNA and DNA. I. Biochem. Biophys. Res. Commun. 11: 107–114.
Gold, M., J. Hurwitz, and M. Anders. 1963. Enzymatic methylation of RNA and DNA. II. On the species specificity of the methylation enzymes. Proc. Nat. Acad. Sci. U. S. A. 50: 164–169.
Meselson, M., and R. Yuan. 1968. DNA restriction enzyme fromE. coli. Nature (London) 217: 1110–1114.
Hattman, S., E. Gold, and A. Plotnik. 1972. Methylation of cytosine residues in DNA controlled by a drug resistance factor. Proc. Nat. Acad. Sci. U. S. A. 69: 187–190.
Scarano, E., M. Iaccarino, P. Grippo, and E. Parisi. 1967. The heterogeneity of thymine methyl group origin in DNA pyrimidine isostichs of developing sea urchin embryos. Proc. Nat. Acad. Sci. U. S. A. 57: 1394–1400.
Lark, C. 1968. Effect of the methionine analogs, ethionine and norleucine, on DNA synthesis inEscherichia coli 15T−. J. Mol. Biol. 31: 401–414.
Maio, J. 1971. DNA strand reassociation and polyribonucleotide binding in the African green monkey,Cercopithecus aethiops. J. Mol. Biol. 56: 579–595.
Ritzi, E., and A. J. Levine. 1969. DNA replication in simian virus 40-infected cells. J. Virol. 5: 686–692.
Gantt, R., and V. J. Evans. 1969. Comparison of soluble RNA methylase capacity in paired neoplastic and nonneoplastic cell linesin vitro. Cancer Res. 29: 536–541.
Evans, V. J., and W. F. Andresen. 1966. Effect of serum on spontaneous neoplastic transformationin vitro. J. Nat. Cancer Inst. 37: 247–249.
Flamm, W., M. Birnstiel, and P. M. B. Walker. 1967. Preparation and fractionation, and isolation of single strands of DNA by isopycnic ultracentrifugation in fixed-angle rotors. In: G. D. Birnie and S. M. Fox (Eds.),Subcellular Components, Preparation and Fractionation. Plenum Press, New York, pp. 125–155.
Srinivasan, P. R., and E. Borek. 1964. Enzymatic alteration of nucleic acid structure. Science 145: 548–553.
Salomon, R., A. M. Kaye, and M. Herzberg. 1969. Mouse nuclear satellite DNA: 5-methyl-cytosine content, pyrimidine isoplith distribution and electron microscopic appearance. J. Mol. Biol. 43: 581–592.
Flamm, W. G., P. M. B. Walker, and M. McCallum. 1969. Some properties of the single strands isolated from the DNA of the nuclear satellite of the mouse (Mus musculus). J. Mol. Biol. 40: 423–443.
Flamm, W. G., M. McCallum, and P. M. B. Walker. 1967. The isolation of complementary strands from a mouse DNA fraction. Proc. Nat. Acad. Sci., U. S. A. 57: 1729–1734.
Smith, B. J. 1970. Light satellite-band DNA in mouse cells infected with polyoma virus. J. Mol. Biol. 47: 101–106.
Flamm, W. G., N. J. Bernheim, and P. E. Brubaker. 1971. Density gradient analysis of newly replicated DNA from synchronized mouse lymphoma cells. Exp. Cell Res. 64: 97–104.
Dawid, I. B., D. D. Brown, and R. H. Reeder. 1970. Composition and structure of chromosomal and amplified ribosomal DNA's ofXenopus laevis. J. Mol. Biol. 51: 341–360.
Sneider, T. W., and V. R. Potter. 1969. Methylation of mammalian DNA: studies on Novikoff hepatoma cells in tissue culture. J. Mol. Biol. 42: 271–284.
Flamm, W. G., N. J. Berheim, and J. Spalding. 1969. Selective inhibition of the semicon-servative replication of mouse satellite DNA. Biochim. Biophys. Acta 195: 273–275.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Gantt, R., De Oca, F.M. & Evans, V.J. Methylation of satellite deoxyribonucleic acid in mouse neoplastic and non-neoplastic cell cultures. In Vitro 8, 288–294 (1973). https://doi.org/10.1007/BF02615908
Issue Date:
DOI: https://doi.org/10.1007/BF02615908