DNA Methylase Covalent Complexes in Cancer

癌症中的 DNA 甲基化酶共价复合物

• 批准号: 7055064
• 负责人: MARK T MULLER
• 金额: $ 23.45万
• 依托单位: UNIVERSITY OF CENTRAL FLORIDA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-02-09 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7055064
• 关键词: CpG islands; DNA methylation; DNA replication; binding sites; bioinformatics; enzyme activity; enzyme substrate complex; genome; methyltransferase; nucleic acid purification; polymerase chain reaction; protein isoforms

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.

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因其失调的后果而凸显，其中包括严重的遗传缺陷和癌症。由于甲基化是由许多不同的酶进行的，因此该过程很复杂。然而，它们在体内的作用却知之甚少。提出了一种了解所有 DNA 甲基转移酶作用的全球性全基因组方法，该方法使用新技术，可以快速查看体内任何给定细胞中的结合事件。这种方法被称为体内甲基酶复合物 (ICM)，使研究人员能够识别染色体环境中几乎所有具有催化活性的内源甲基化酶的基因和同源 DNA 结合位点。由于 ICM 测定是基于抗体的，因此可以轻松区分甲基转移酶亚型。该方法是高度定量且易于处理的。基于当前 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，以检查活细胞中甲基化酶与基因组相互作用的微妙之处。鉴于甲基化作为分子靶标的强烈兴趣，这些实验也与使用 DNA 低甲基化剂开发有效的抗癌疗法相关。最后，ICM 方法代表了一种明确定义的新型生物学相关性，可能对 DNA 低甲基化药物的临床试验带来显着益处。