Isolation and characterization of a novel zinc finger protein MIZF in DNA methylation and transcriptional regulation

新型锌指蛋白 MIZF 在 DNA 甲基化和转录调控中的分离和表征

• 批准号: 13680720
• 负责人: SEKIMATA Masayuki
• 金额: $ 2.3万
• 依托单位: Fukushima Medical University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13680720/
• 关键词: DNA methylation; Trascription factor; Zinc finger; Methyl CpG-binding protein; ジンクフィンガー分子; クロマチン; ヒストン修飾

MBD2 is a member of the methyl-CpG binding protein family that plays an important role in methylated DNA silencing. We have recently identified a novel zinc finger protein, MIZF, as an MBD2-binding partner. To understand the physiological function of MIZF in MBD2-mediated gene silencing, we investigated the DNA-binding properties of MIZF and its potential target genes. Using a cyclic amplification and selection of targets technique, the DNA sequence CGGACGTT was determined as sufficient for MIZF binding. Deletion of individual zinc fingers revealed that five of the seven zinc fingers are required for DNA binding. Reporter assays demonstrated that MIZF represses transcription from the promoter including this DNA sequence. Database search indicated that a variety of human genes including Rb contain this sequence in their promoter region. MIZE actually bound to its recognition sequence within the Rb promoter and repressed the Rb transcription. These results suggest that MIZF, through its DNA-binding activity, acts as a sequence-specific transcriptional repressor likely involved in MBD2-mediated epigenetic genesilencing.

MBD 2是甲基-CpG结合蛋白家族的成员，其在甲基化DNA沉默中起重要作用。我们最近发现了一种新的锌指蛋白，MIZF，作为MBD 2结合伙伴。为了了解MIZF在MBD 2介导的基因沉默中的生理功能，我们研究了MIZF及其潜在靶基因的DNA结合特性。使用循环扩增和靶点选择技术，确定DNA序列CGGACGTT足以与MIZF结合。个别锌指的缺失揭示了七个锌指中的五个是DNA结合所需的。报告基因分析表明，MIZF抑制从启动子包括这个DNA序列的转录。数据库检索表明，包括Rb在内的多种人类基因在其启动子区含有该序列。米泽实际上与Rb启动子内的识别序列结合并抑制Rb转录。这些结果表明，MIZF，通过其DNA结合活性，作为一个序列特异性转录抑制可能参与MBD 2介导的表观遗传基因沉默。

项目成果

SEKIMATA Masayuki: "Involvement of a Novel Zinc Finger Protein, MIZF, in Transcriptional Repression by Interacting with a Methyl-CpG-binding Protein, MBD2"J.Biol.Chem.. 276(46). 42632-42638 (2001)

SEKIMATA Masayuki：“新型锌指蛋白 MIZF 通过与甲基 CpG 结合蛋白 MBD2 相互作用参与转录抑制”J.Biol.Chem. 276(46)。

KABUYAMA Yukihiko: "Wavelength-specific activation of MAP kinase family proteins by monochromatic UV irradiation"Photochem.Photobiol. 73(2). 147-152 (2001)

KABUYAMA Yukihiko：“单色 UV 照射对 MAP 激酶家族蛋白的波长特异性激活”Photochem.Photobiol。

Sekimata Masayuki: "Involvement of a novel zinc finger protein, MIZF, in transcriptional repression by interacting with a methyl-CpG binding protein, MBD2"Journal of Biological Chemistry. 276・46. 42632-42638 (2001)

Sekimata Masayuki：“新型锌指蛋白 MIZF 通过与甲基 CpG 结合蛋白 MBD2 相互作用参与转录抑制”生物化学杂志 276・46 (2001)。

SEKIMATA Masayuki: "Sequence-Specific Transcriptional Repression by an MBD2-Interacting Zinc Finger Protein MIZE"Nucleic Acids Res.. 32. 590-597 (2004)

SEKIMATA Masayuki：“MBD2 相互作用的锌指蛋白 MIZE 的序列特异性转录抑制”核酸研究.. 32. 590-597 (2004)

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