Abstract
Monoclonal antibodies prepared against DNA methyltransferase from human placenta undergo immune complex formation also with DNA methyltransferase from P815 mouse mastocytoma cells. One of these monoclonal antibodies, M2B10, was used for the immunoaffinity purification of this enzyme. Complexes of the immunoaffinity-purified mouse DNA methyltransferase with DNA were visualized by electron microscopy. DNA methyltransferase was found to be distributed along linearized plasmid DNA with a higher incidence of enzyme molecules at the terminal segments. This binding to strand ends was significantly increased after dG- or dGdC-tailing of the DNA, which is compatible with a preferred binding of the enzyme to single-stranded DNA. Sequence specificity analysis using methyl-sensitive restriction enzymes showed that the mouse DNA methyltransferase transferred methyl groups to the internal cytosines in 5'CCGG and 5'GCGC sequences, however, the external cytosine in 5'CCGG sequences was also methylated.
Full text
PDF





Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Adams R. L., McKay E. L., Craig L. M., Burdon R. H. Mouse DNA methylase: methylation of native DNA. Biochim Biophys Acta. 1979 Feb 27;561(2):345–357. doi: 10.1016/0005-2787(79)90143-6. [DOI] [PubMed] [Google Scholar]
- Bestor T. H., Ingram V. M. Growth-dependent expression of multiple species of DNA methyltransferase in murine erythroleukemia cells. Proc Natl Acad Sci U S A. 1985 May;82(9):2674–2678. doi: 10.1073/pnas.82.9.2674. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bestor T. H., Ingram V. M. Two DNA methyltransferases from murine erythroleukemia cells: purification, sequence specificity, and mode of interaction with DNA. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5559–5563. doi: 10.1073/pnas.80.18.5559. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bolden A., Ward C., Siedlecki J. A., Weissbach A. DNA methylation. Inhibition of de novo and maintenance methylation in vitro by RNA and synthetic polynucleotides. J Biol Chem. 1984 Oct 25;259(20):12437–12443. [PubMed] [Google Scholar]
- Busslinger M., deBoer E., Wright S., Grosveld F. G., Flavell R. A. The sequence GGCmCGG is resistant to MspI cleavage. Nucleic Acids Res. 1983 Jun 11;11(11):3559–3569. doi: 10.1093/nar/11.11.3559. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Doerfler W. DNA methylation and gene activity. Annu Rev Biochem. 1983;52:93–124. doi: 10.1146/annurev.bi.52.070183.000521. [DOI] [PubMed] [Google Scholar]
- Drahovsky D., Morris N. R. The mechanism of action of rat liver DNA methylase. 3. Nucleotide requirements for binding and methylation. Biochim Biophys Acta. 1972 Aug 25;277(2):245–250. doi: 10.1016/0005-2787(72)90404-2. [DOI] [PubMed] [Google Scholar]
- Drahovský D., Morris N. R. Mechanism of action of rat liver DNA methylase. I. Interaction with double-stranded methyl-acceptor DNA. J Mol Biol. 1971 May 14;57(3):475–489. doi: 10.1016/0022-2836(71)90104-5. [DOI] [PubMed] [Google Scholar]
- Drahovský D., Morris N. R. Mechanism of action of rat liver DNA methylase. II. Interaction with single-stranded methyl-acceptor DNA. J Mol Biol. 1971 Oct 28;61(2):343–356. doi: 10.1016/0022-2836(71)90384-6. [DOI] [PubMed] [Google Scholar]
- Grafstrom R. H., Yuan R., Hamilton D. L. The characteristics of DNA methylation in an in vitro DNA synthesizing system from mouse fibroblasts. Nucleic Acids Res. 1985 Apr 25;13(8):2827–2842. doi: 10.1093/nar/13.8.2827. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gruenbaum Y., Cedar H., Razin A. Substrate and sequence specificity of a eukaryotic DNA methylase. Nature. 1982 Feb 18;295(5850):620–622. doi: 10.1038/295620a0. [DOI] [PubMed] [Google Scholar]
- Gruenbaum Y., Szyf M., Cedar H., Razin A. Methylation of replicating and post-replicated mouse L-cell DNA. Proc Natl Acad Sci U S A. 1983 Aug;80(16):4919–4921. doi: 10.1073/pnas.80.16.4919. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grünwald S., Drahovsky D. DNA-cytosine-5-methyltransferase from P815 mouse mastocytoma cells: "maintenance" and "de novo" activities are carried out by the same enzyme molecule. Int J Biochem. 1984;16(8):883–888. doi: 10.1016/0020-711x(84)90147-2. [DOI] [PubMed] [Google Scholar]
- Jaenisch R., Jähner D. Methylation, expression and chromosomal position of genes in mammals. Biochim Biophys Acta. 1984 May 15;782(1):1–9. doi: 10.1016/0167-4781(84)90099-x. [DOI] [PubMed] [Google Scholar]
- Jones P. A., Taylor S. M. Hemimethylated duplex DNAs prepared from 5-azacytidine-treated cells. Nucleic Acids Res. 1981 Jun 25;9(12):2933–2947. doi: 10.1093/nar/9.12.2933. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kaul S., Pfeifer G. P., Drahovsky D. Preparation of monoclonal antibodies against DNA-cytosine-5-methyltransferase from human placenta. Eur J Cell Biol. 1984 Jul;34(2):330–335. [PubMed] [Google Scholar]
- Keshet E., Cedar H. Effect of CpG methylation on Msp I. Nucleic Acids Res. 1983 Jun 11;11(11):3571–3580. doi: 10.1093/nar/11.11.3571. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- McClelland M. The effect of site specific methylation on restriction endonuclease cleavage (update). Nucleic Acids Res. 1983 Jan 11;11(1):r169–r173. doi: 10.1093/nar/11.1.235-c. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Merril C. R., Goldman D., Sedman S. A., Ebert M. H. Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science. 1981 Mar 27;211(4489):1437–1438. doi: 10.1126/science.6162199. [DOI] [PubMed] [Google Scholar]
- Pfeifer G. P., Grünwald S., Boehm T. L., Drahovsky D. Isolation and characterization of DNA cytosine 5-methyltransferase from human placenta. Biochim Biophys Acta. 1983 Aug 2;740(3):323–330. doi: 10.1016/0167-4781(83)90141-0. [DOI] [PubMed] [Google Scholar]
- Razin A., Szyf M. DNA methylation patterns. Formation and function. Biochim Biophys Acta. 1984 Sep 10;782(4):331–342. doi: 10.1016/0167-4781(84)90043-5. [DOI] [PubMed] [Google Scholar]
- Salomon R., Kaye A. M. Methylation of mouse DNA in vivo: di- and tripyrimidine sequences containing 5-methylcytosine. Biochim Biophys Acta. 1970 Apr 15;204(2):340–351. [PubMed] [Google Scholar]
- Sneider T. W., Teague W. M., Rogachevsky L. M. S-adenosylmethionine: DNA-cytosine 5-methyltransferase from a Novikoff rat hepatoma cell line. Nucleic Acids Res. 1975 Oct;2(10):1685–1700. doi: 10.1093/nar/2.10.1685. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sneider T. W. The 5'-cytosine in CCGG1 is methylated in two eukaryotic DNAs and Msp I is sensitive to methylation at this site. Nucleic Acids Res. 1980 Sep 11;8(17):3829–3840. doi: 10.1093/nar/8.17.3829. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sutcliffe J. G. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. doi: 10.1101/sqb.1979.043.01.013. [DOI] [PubMed] [Google Scholar]
- Taylor S. M., Jones P. A. Mechanism of action of eukaryotic DNA methyltransferase. Use of 5-azacytosine-containing DNA. J Mol Biol. 1982 Dec 15;162(3):679–692. doi: 10.1016/0022-2836(82)90395-3. [DOI] [PubMed] [Google Scholar]
- Wang R. Y., Huang L. H., Ehrlich M. Human placental DNA methyltransferase: DNA substrate and DNA binding specificity. Nucleic Acids Res. 1984 Apr 25;12(8):3473–3490. doi: 10.1093/nar/12.8.3473. [DOI] [PMC free article] [PubMed] [Google Scholar]



