Research on the regulation of vertebrates' genome DNA methylation

脊椎动物基因组DNA甲基化调控研究

• 批准号: 17370047
• 负责人: TAJIMA Shoji
• 金额: $ 10.05万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17370047/
• 关键词: DNA methylation; DNA methvltransferase; nucleosome; hisotne; Dnmtl; Dnmt3a; Dnmt3b; Np95; Dnmt3L; PCNA; 遺伝情報発現制御

Genome in higher eukaryotes suffers methylation modification. This methylation affects not the coding but the expression information. The methylation modification is catalyzed by three DNA methyltransferases (Dnmt) and one related factor Dnmt3L, which has no catalytic activity. In the research project, we aimed to elucidate the molecular mechanism of the Dnmt on the creation and the maintenance of DNA methylation patterns, and obtained the following results.Dnmtl, which contributes to the maintenance of genome methylation patterns, processively methylates hemimethylated CpG on a double-stranded DNA. Unexpectedly, Dnmt1 frequently fails to methylate hemimethylated positions. The N-terminal portion of Dnmt1 forms independent domain structure, and possesses DNA binding activity that recognizes minor grove of DNA. We found Np95 as a new factor that is necessary for the maintenance of DNA methylation patterns in vivo. These properties help in understanding the mechanism of the maintenance of DNA methylation patterns.De novo-type enzyme Dnmt3a hardly methylates nucleosome core region, but that Dnmt3b can significantly methylates. Dnmt3a efficiently methylates the naked linker region of nucleosomes, and the activity is inhibited by linker histone H1. Histone H1 may contribute to the regulation in creating DNA methylation patterns by Dnmt3a. In male primordial germ cells (PGC), in which global methylation occurs, Dnmt3a2, which is an isoform of Dnmt3a, and Dnmt3L are highly expressed. Dnmt3L is prerequisite for the global methylation. We found that Dnmt3a2 cannot show catalytic activity under physiological salt conditions, but restores the activity in the presence of Dnmt3L. It is suggested that this salt-dependent catalytic activity of Dnmt3a2 is the reason why the global methylation in PGC requires Dnmt3L. These properties of Dnmt3 help in understanding the creation of DNA methylation patterns.

高等真核生物基因组发生甲基化修饰。这种甲基化不影响编码，而是影响表达信息。甲基化修饰由三种DNA甲基转移酶（Dnmt）和一种无催化活性的相关因子Dnmt 3L催化。在本研究项目中，我们旨在阐明Dnmt在DNA甲基化模式的产生和维持中的分子机制，并获得了以下结果：Dnmtl有助于基因组甲基化模式的维持，它使双链DNA上的半甲基化CpG原甲基化。出乎意料的是，Dnmt 1经常不能使半甲基化位置甲基化。Dnmt 1的N端部分形成独立的结构域结构，并具有识别DNA小格罗夫的DNA结合活性。我们发现Np95是一种新因子，对于体内维持DNA甲基化模式是必要的。这些性质有助于理解DNA甲基化模式维持的机制，新类型的酶Dnmt 3a几乎不甲基化核小体核心区，但Dnmt 3b可以显著甲基化。Dnmt3a有效地甲基化核小体的裸露接头区域，并且该活性被接头组蛋白H1抑制。组蛋白H1可能参与Dnmt3a对DNA甲基化模式的调控。在发生整体甲基化的雄性原始生殖细胞（PGC）中，作为Dnmt3a的同种型的Dnmt3a2和Dnmt3L高度表达。Dnmt3L是全局甲基化的先决条件。我们发现Dnmt3a2在生理盐条件下不能显示催化活性，但在Dnmt3L存在下恢复活性。这表明Dnmt3a2的这种盐依赖性催化活性是PGC中的全局甲基化需要Dnmt3L的原因。Dnmt3的这些特性有助于理解DNA甲基化模式的产生。

项目成果

Bmi1 cooperates with Dnmt1-associated protein 1 in gene silencing

• DOI: 10.1016/j.bbrc.2006.12.166
• 发表时间: 2007-02-23
• 期刊: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
• 影响因子: 3.1
• 作者: Negishi, Masamitsu;Saraya, Atsunori;Iwama, Atsushi
• 通讯作者: Iwama, Atsushi

Mechanism of maintenance and formation of DNA methylation patterns

DNA甲基化模式的维持和形成机制

• 发表时间: 2005
• 作者: Shoji;Tajima
• 通讯作者: Tajima

Cyclin-dependent kinase like 5 (CDKL5) binds to the N-terminal region of DNA methyltransferase 1 (Dnmt1)

细胞周期蛋白依赖性激酶样 5 (CDKL5) 与 DNA 甲基转移酶 1 (Dnmt1) 的 N 末端区域结合

• 发表时间: 2007
• 作者: Daisuke;Hamasaki
• 通讯作者: Hamasaki

DNAメチルトランスフェラーゼ1(Dnmt1)のN末端領域に結合するプロテインキナーゼの同定と解析

与 DNA 甲基转移酶 1 (Dnmt1) N 末端区域结合的蛋白激酶的鉴定和分析

• 发表时间: 2007
• 作者: Ohno-Jinno A;Isogai Z;Yoneda M;Kasai K;Miyaishi O;Inoue Y;Kataoka T;Zhao JS;Li H;Takeyama M;Keene DR;Sakai LY;Kimata K;Iwaki M;Zako M;Hikosaka K;関口 茉里
• 通讯作者: 関口 茉里

ポリコーム遺伝子Bmi-1によるDNAメチル化パターンの維持とその分子機構

多梳基因Bmi-1维持DNA甲基化模式及其分子机制

• 发表时间: 2005
• 作者: Garantziotis S;Hollingsworth JW;Ghanayem RB;Timberlake S;Zhuo L;Kimata K;Schwartz DA.;根岸 正充
• 通讯作者: 根岸 正充