A column method for determination of DNA cytosine-C5-methyltransferase activity

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A column method for determination of DNA cytosine-C5-methyltransferase activity
Bo Yeon Kim; Oh Sung Kwon; Sung Ah Joo; Jung Ah Park; Gun Young Heo; Min Soo Kim; Jong Seog Ahn
Bibliographic Citation
Analytical Biochemistry, vol. 326, no. 1, pp. 21-24
Publication Year
DNA methylation at the 5th position of cytosine has been found to be correlated with tumorigenesis. An inhibitor of DNA methylase could, therefore, be used as an anticancer drug. However, only a few inhibitory compounds have been discovered due to the limitations for assaying the DNA methylation. In this study, we describe a modification of DNA cytosine-C5-methyltransferase assay system utilizing [3H]-labeled S-adenosyl-methionine (SAM) and Sephadex G-25 column. Pre-treatment of either λ DNA or the promoter region of human telomerase (hTERT) with HaeIII methylase greatly reduced the digestion of the DNAs with the corresponding restriction enzyme HaeIII endonuclease (over 100-fold), and the result was further confirmed by agarose gel electrophoresis. Application of this column method to another modification/restriction system, EcoRI methylase/endonuclease, gave rise to the similar results. Our data suggest that the newly developed column method could be effective for rapid screening of large number of cytosine methylase inhibitors and could also be applicable to other DNA methylases.
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Ochang Branch Institute > Chemical Biology Research Center > 1. Journal Articles
Jeonbuk Branch Institute > Microbial Biotechnology Research Center > 1. Journal Articles
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