DNA methylation is an epigenetic encryption system that is essential for life. The importance of DNA methylation in man is underscored by the consequences of its misregulation, which include and serious genetics defects and cancer. The process is complicated by the fact that methylation is carried out by a number of different enzymes; however their in vivo roles are poorly understood. A global, genome wide approach to understanding the action of all DNA methyltransferases is proposed using novel technology that allows a snapshot view of the binding event in any given cell in vivo. This method, called the In vivo Complex of Methylase (ICM), will allow empower researchers with the ability to identify genes and cognate DNA binding sites for virtually all catalytically active, endogenous methylases in a chromosomal setting. Because the ICM assay is antibody based, one can readily differentiate methyltransferase isoforms. The approach is highly quantitative and tractable. A series of experiments are proposed, based on current tools of biochemistry and molecular biology of DNA methylation reaction mechanism, to examine subtleties of methylase:genome interactions in living cells. Given the acute interest of methylation as a molecular target, these experiments are also relevant to the development of effective anti-cancer therapy using DNA hypomethylating agents. Finally, the ICM method represents a clearly defined and novel biological correlate that could be a significant benefit in clinical trials with DNA hypomethylating drugs. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
7R01CA098214-03
Application #
7055064
Study Section
Alcohol and Toxicology Subcommittee 4 (ALTX)
Program Officer
Knowlton, John R
Project Start
2004-02-09
Project End
2008-01-31
Budget Start
2005-02-03
Budget End
2006-01-31
Support Year
3
Fiscal Year
2005
Total Cost
$234,520
Indirect Cost
Name
University of Central Florida
Department
Biochemistry
Type
Schools of Medicine
DUNS #
150805653
City
Orlando
State
FL
Country
United States
Zip Code
32826
Lee, Bongyong; Morano, Annalisa; Porcellini, Antonio et al. (2012) GADD45? inhibition of DNMT1 dependent DNA methylation during homology directed DNA repair. Nucleic Acids Res 40:2481-93
Cyril, Vidusha; Muller, Mark T (2012) A solid phase assay for topoisomerase I interfacial poisons and catalytic inhibitors. Anal Biochem 421:607-16
Lee, Gun E; Kim, Joo Hee; Taylor, Michael et al. (2010) DNA methyltransferase 1-associated protein (DMAP1) is a co-repressor that stimulates DNA methylation globally and locally at sites of double strand break repair. J Biol Chem 285:37630-40
Lee, Bongyong; Muller, Mark T (2009) SUMOylation enhances DNA methyltransferase 1 activity. Biochem J 421:449-61
Lee, Gun Eui; Yu, Eun Young; Cho, Chae Hyun et al. (2004) DNA-protein kinase catalytic subunit-interacting protein KIP binds telomerase by interacting with human telomerase reverse transcriptase. J Biol Chem 279:34750-5