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Title 

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

Authors 

Bo Yeon KimOh Sung KwonSung Ah JooJung Ah ParkGun Young HeoMin Soo KimJong Seog Ahn

Publisher 

Elsevier

Issue Date 

2004

Citation 

Analytical Biochemistry, vol. 326, no. 1, pp. 21-24

Keywords 

DNA (cytosine 5) methyltransferaseexodeoxyribonuclease IIIs adenosylmethioninesephadextelomeraseagar gel electrophoresisanalytic methodDNA degradationDNA modificationDNA screening

Abstract 

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.

ISSN 

0003-2697

Link 

http://dx.doi.org/10.1016/j.ab.2003.11.009

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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