BIOMAT Database Logo BIOMAT Database Logo

A column method for determination of DNA cytosine-C5-methyltransferase activity. 2004

Bo Yeon Kim, and Oh Sung Kwon, and Sung Ah Joo, and Jung Ah Park, and Keon Young Heo, and Min Soo Kim, and Jong Seog Ahn
Laboratory of Cellular Signaling Modulator, Korea Research Institute of Bioscience and Biotechnology, 305-333 Yuseong, Daejon, South Korea.

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 [(3)H]-labeled S-adenosyl-methionine (SAM) and Sephadex G-25 column. Pre-treatment of either lambda 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.

UI MeSH Term Description Entries
D010582 Bacteriophage lambda A temperate inducible phage and type species of the genus lambda-like viruses, in the family SIPHOVIRIDAE. Its natural host is E. coli K12. Its VIRION contains linear double-stranded DNA with single-stranded 12-base 5' sticky ends. The DNA circularizes on infection. Coliphage lambda,Enterobacteria phage lambda,Phage lambda,lambda Phage
D011401 Promoter Regions, Genetic DNA sequences which are recognized (directly or indirectly) and bound by a DNA-dependent RNA polymerase during the initiation of transcription. Highly conserved sequences within the promoter include the Pribnow box in bacteria and the TATA BOX in eukaryotes. rRNA Promoter,Early Promoters, Genetic,Late Promoters, Genetic,Middle Promoters, Genetic,Promoter Regions,Promoter, Genetic,Promotor Regions,Promotor, Genetic,Pseudopromoter, Genetic,Early Promoter, Genetic,Genetic Late Promoter,Genetic Middle Promoters,Genetic Promoter,Genetic Promoter Region,Genetic Promoter Regions,Genetic Promoters,Genetic Promotor,Genetic Promotors,Genetic Pseudopromoter,Genetic Pseudopromoters,Late Promoter, Genetic,Middle Promoter, Genetic,Promoter Region,Promoter Region, Genetic,Promoter, Genetic Early,Promoter, rRNA,Promoters, Genetic,Promoters, Genetic Middle,Promoters, rRNA,Promotor Region,Promotors, Genetic,Pseudopromoters, Genetic,Region, Genetic Promoter,Region, Promoter,Region, Promotor,Regions, Genetic Promoter,Regions, Promoter,Regions, Promotor,rRNA Promoters
D003911 Dextrans A group of glucose polymers made by certain bacteria. Dextrans are used therapeutically as plasma volume expanders and anticoagulants. They are also commonly used in biological experimentation and in industry for a wide variety of purposes. Dextran,Dextran 40,Dextran 40000,Dextran 70,Dextran 75,Dextran 80,Dextran B-1355,Dextran B-1355-S,Dextran B1355,Dextran B512,Dextran Derivatives,Dextran M 70,Dextran T 70,Dextran T-40,Dextran T-500,Hemodex,Hyskon,Infukoll,Macrodex,Polyglucin,Promit,Rheodextran,Rheoisodex,Rheomacrodex,Rheopolyglucin,Rondex,Saviosol,Dextran B 1355,Dextran B 1355 S,Dextran T 40,Dextran T 500
D004248 DNA (Cytosine-5-)-Methyltransferases Enzymes that catalyzes the transfer of a methyl group from S-ADENOSYLMETHIONINE to the 5-position of CYTOSINE residues in DNA. DNA (Cytosine-5-)-Methyltransferase,DNA Cytosine-5-Methylase,DNA (Cytosine 5) Methyltransferase,Cytosine-5-Methylase, DNA,DNA Cytosine 5 Methylase
D004269 DNA, Bacterial Deoxyribonucleic acid that makes up the genetic material of bacteria. Bacterial DNA
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D019098 Telomerase An essential ribonucleoprotein reverse transcriptase that adds telomeric DNA to the ends of eukaryotic CHROMOSOMES. Telomerase Catalytic Subunit,Telomerase Reverse Transcriptase,Telomerase Reverse Transcriptase Catalytic Subunit,Catalytic Subunit, Telomerase,Reverse Transcriptase, Telomerase,Subunit, Telomerase Catalytic,Transcriptase, Telomerase Reverse
D019175 DNA Methylation Addition of methyl groups to DNA. DNA methyltransferases (DNA methylases) perform this reaction using S-ADENOSYLMETHIONINE as the methyl group donor. DNA Methylations,Methylation, DNA,Methylations, DNA

Related Publications

Bo Yeon Kim, and Oh Sung Kwon, and Sung Ah Joo, and Jung Ah Park, and Keon Young Heo, and Min Soo Kim, and Jong Seog Ahn
The human Dnmt2 has residual DNA-(cytosine-C5) methyltransferase activity.
August 2003, The Journal of biological chemistry,
Bo Yeon Kim, and Oh Sung Kwon, and Sung Ah Joo, and Jung Ah Park, and Keon Young Heo, and Min Soo Kim, and Jong Seog Ahn
DNA binding discrimination of the murine DNA cytosine-C5 methyltransferase.
May 1998, Journal of molecular biology,
Bo Yeon Kim, and Oh Sung Kwon, and Sung Ah Joo, and Jung Ah Park, and Keon Young Heo, and Min Soo Kim, and Jong Seog Ahn
A real-time assay for CpG-specific cytosine-C5 methyltransferase activity.
May 2010, Nucleic acids research,
Bo Yeon Kim, and Oh Sung Kwon, and Sung Ah Joo, and Jung Ah Park, and Keon Young Heo, and Min Soo Kim, and Jong Seog Ahn
A potent cell-active allosteric inhibitor of murine DNA cytosine C5 methyltransferase.
March 2003, The Journal of biological chemistry,
Bo Yeon Kim, and Oh Sung Kwon, and Sung Ah Joo, and Jung Ah Park, and Keon Young Heo, and Min Soo Kim, and Jong Seog Ahn
DNA cytosine C5 methyltransferase Dnmt1: catalysis-dependent release of allosteric inhibition.
July 2005, Biochemistry,
Bo Yeon Kim, and Oh Sung Kwon, and Sung Ah Joo, and Jung Ah Park, and Keon Young Heo, and Min Soo Kim, and Jong Seog Ahn
M.BssHII, a multispecific cytosine-C5-DNA-methyltransferase with unusual target recognizing properties.
April 1996, Journal of molecular biology,
Bo Yeon Kim, and Oh Sung Kwon, and Sung Ah Joo, and Jung Ah Park, and Keon Young Heo, and Min Soo Kim, and Jong Seog Ahn
AdoMet-dependent methyl-transfer: Glu119 is essential for DNA C5-cytosine methyltransferase M.HhaI.
November 2007, Journal of molecular biology,
Bo Yeon Kim, and Oh Sung Kwon, and Sung Ah Joo, and Jung Ah Park, and Keon Young Heo, and Min Soo Kim, and Jong Seog Ahn
A 7-Deazaadenosylaziridine Cofactor for Sequence-Specific Labeling of DNA by the DNA Cytosine-C5 Methyltransferase M.HhaI.
November 2015, Molecules (Basel, Switzerland),
Bo Yeon Kim, and Oh Sung Kwon, and Sung Ah Joo, and Jung Ah Park, and Keon Young Heo, and Min Soo Kim, and Jong Seog Ahn
The Dnmt1 DNA-(cytosine-C5)-methyltransferase methylates DNA processively with high preference for hemimethylated target sites.
November 2004, The Journal of biological chemistry,
Bo Yeon Kim, and Oh Sung Kwon, and Sung Ah Joo, and Jung Ah Park, and Keon Young Heo, and Min Soo Kim, and Jong Seog Ahn
An in vitro screening method for DNA cytosine-C5-methylase inhibitor.
February 2002, Natural product letters,
Copied contents to your clipboard!