Structural basis for regulation of chromatin structure by DNA methylation

DNA甲基化调节染色质结构的结构基础

• 批准号: 13480230
• 负责人: SHIRAKAWA Masahiro
• 金额: $ 8.19万
• 依托单位: Yokohama City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13480230/
• 关键词: DNA methylation; ubiquitination; DNA repair; NMR; クロマチン; 動的構造; ヒストン メチル; メチル化DNA結合蛋白質; クロマチン構造; エピジェネティックス; 転写; ヒストン メチル化; メチル化DNA; メチル化DNA結合ドメイン; 立体構造; ミスマッチDNA

The purpose of this study was to clarify the regulation of chromatin structures mediated by genome DNA methylation, and to investigate post-translational modifications of nuclear proteins by ubiquitin-like modifier proteins. We proceeded structural and functional analyses of the methylated DNA binding domain (MBD) of MBD1, the ubiquitin-like domain of human HR23B, and the ubiquitin-associated (UBA) domain of Saccharomyces cerevisae Rad23 and Lys 48- or Lys 63-linked di- or tetra-ubiquitin.We analyzed the dynamic property of the complex between MBD1 MBD and a methylated DNA by using NMR, SPR and isothermal calorimetry. We have determined solution structure of ubiquitin-like domain of human HR23B in complex with a ubiquitin interacting motif of proteasomal S5a subunit by NMR. We also determined solution structure of the complex between the UBA domain of yeast DSK2 and ubiquitin. In addition, we showed that show the difference in structural properties of Lys 48- or Lys 63-linked ubiquitin chains by isotope-aided NMR and small angle X-ray scattering.

本研究的目的是阐明基因组DNA甲基化介导的染色质结构的调节，并研究泛素样修饰蛋白对核蛋白的翻译后修饰。我们对MBD 1的甲基化DNA结合结构域（MBD）、人HR 23 B的泛素样结构域（ubiquitin-like domain）、酿酒酵母Rad 23的泛素相关（乌巴）结构域和赖氨酸48或赖氨酸63连接的二或四泛素结构域进行了结构和功能分析，并利用NMR、SPR和等温量热法分析了MBD 1 MBD与甲基化DNA复合物的动力学性质。我们用核磁共振方法确定了人HR 23 B的泛素样结构域与蛋白酶体S5 a亚基的泛素相互作用基序复合物的溶液结构。我们还测定了酵母DSK 2的乌巴结构域与泛素复合物的溶液结构。此外，我们还通过同位素辅助的核磁共振和小角X射线散射研究了Lys 48-和Lys 63-连接的泛素链在结构性质上的差异。

项目成果

Jee, J.-G., Ikegami, T., Hashimoto, M.他: "Solution Structure of the Fibronection TypeIII Domain from Bacillus cirulans WL-12 Chitinose A1"Journal of Biological Chemistry. 277. 1388-1397 (2002)

Jee, J.-G.、Ikegami, T.、Hashimoto, M. 等人：“来自卷状芽孢杆菌 WL-12 几丁糖 A1 的纤维连接 III 型结构域的溶液结构”《生物化学杂志》277. 1388-1397 (2002)。 ）

Kouno, T., Miura, K., Kanematsu, T.他: "1H, 13C and 15N resonance assignments of GABARAP, GABAA receptor associated protein"J. biomol. NMR. (印刷中).

Kouno, T.、Miura, K.、Kanematsu, T. 等人：“GABARAP、GABAA 受体相关蛋白的 1H、13C 和 15N 共振分配”J. Biomol。

The structure and binding mode of interleukin-18

• DOI: 10.1038/nsb993
• 发表时间: 2003-11-01
• 期刊: NATURE STRUCTURAL BIOLOGY
• 作者: Kato, Z;Jee, J;Shirakawa, M
• 通讯作者: Shirakawa, M

Kouno, T., Miura, K., Kanematsu, T., Shirakawa, M., Hirata, M., 他: "1H, 13C and 15N resonance assignments of GABARAP, GABAA receptor associated protein"Journal of Biomolecular NMR. 22. 97-98 (2002)

Kouno, T.、Miura, K.、Kanematsu, T.、Shirakawa, M.、Hirata, M.等人：“GABARAP、GABAA 受体相关蛋白的 1H、13C 和 15N 共振分配”《生物分子 NMR 杂志》。 22. 97-98 (2002)

Fujiwara, K., Tenno, T., Sugasawa, K., Jee, J.-G., Ohki, I., Kojima, C., Tochio, H., Hiroaki, H., Hanaoka, F., Shirakawa, M.: "Structure of the ubiquitin-interacting motif of S5a bound to the ubiquitin-like domain of HR23B"Journal of Biological Chemistry.

Fujiwara, K.、Tenno, T.、Sugasawa, K.、Jee, J.-G.、Ohki, I.、Kojima, C.、Tochio, H.、Hiroaki, H.、Hanaoka, F.、Shirakawa,