DNA Methylase Covalent Complexes in Cancer

描述（由申请人提供）：DNA甲基化是一种对生命至关重要的表观遗传加密系统。DNA甲基化在人类中的重要性被其失调的后果所强调，其中包括严重的遗传缺陷和癌症。这个过程是复杂的，甲基化是由许多不同的酶进行的;然而，它们在体内的作用知之甚少。一个全球性的，全基因组的方法来了解所有的DNA甲基转移酶的行动，提出了使用新的技术，允许在任何给定的细胞在体内的结合事件的快照视图。这种方法被称为甲基化酶体内复合物（ICM），将使研究人员能够识别染色体环境中几乎所有催化活性内源性甲基化酶的基因和同源DNA结合位点。由于ICM检测是基于抗体的，因此可以容易地区分甲基转移酶亚型。该方法是高度定量和易处理的。基于DNA甲基化反应机制的生物化学和分子生物学工具，提出了一系列实验来研究活细胞中甲基化酶与基因组相互作用的微妙之处。考虑到甲基化作为分子靶点的迫切兴趣，这些实验也与使用DNA低甲基化剂的有效抗癌疗法的开发相关。最后，ICM方法代表了一种明确定义的新生物学相关性，可能在DNA低甲基化药物的临床试验中具有显著益处。

DESCRIPTION (provided by applicant): 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.