STUDIES ON TRANSCRIPTION PROGRESSION FACTORS AND THE APPLICATION

转录进展因子的研究及应用

• 批准号: 03660096
• 负责人: YOSHIDA Michiteru
• 金额: $ 1.28万
• 依托单位: SCIENCE UNIVERSITY OF TOKYO, DEPARTMENT OF BIOLOGICAL SCIENCE AND TECHNOLOGY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03660096/
• 关键词: HMG PROTEINS; HMG BOX; DNA-BINDING PROTEINS; TRANSCRIPTION PROGRESSION; REGULATION OF GENE EXPRESSION; DNA-PROTEIN INTERACTION; HMG box; DNA‐タンパク質相互作用; 核タンパク質; 非ヒストンタンパク質; DNA結合構造; 遺伝子発現調節; 細胞増殖; 外来遺伝子の高発現

(1) The binding of HMG1 and HMG2 with DNA : The binding of HMG1/2 proteins with plasmid DNA of various conformations was examined. The results indicated that HMG1/2 preferentially bind to negatively supercoiled DNA recognizing the conformation without sequence spcificity, and change the DNA topology. The cooperative binding of HMG1/2 to the unusual-structured DNA under torsional stress may, thus, remove the transcriptional block by dissolving the structure. The two DNA binding domains in the HMG protein molecule were over-expressed in E. coil system. The tertiary structures of the purified domains are under investigation.(2) Expression of HNG1 and HMG2 genes dependent on the cell cycle and growth : The expression levels of HMG1/2 genes during cell cycle and growth were analyzed. The results showed that HMG2 gene expression is enhanced at G2 phase in the cell cycle and depending on the cell proliferation, in contrast with HMG1 gene. The relative amount of HMG proteins in the nuclei correlated with the expression level of the respective genes.(3) HMG1 protein stimulates in vivo transcription : The effect of HMG1 gene expression in cells co-transfected with a reporter gene was investigated. The results showed that HMG1 protein stimulates the transcriptional activity of the gene co-transfected. The in vivo system may be useful for the effective production of functionally-active proteins.

(1) HMG1和HMG2与DNA的结合：检测HMG1/2蛋白与不同构象的质粒DNA的结合。结果表明，HMG1/2优先与负超螺旋DNA结合，识别非序列特异性的构象，并改变DNA拓扑结构。因此，在扭转应力下，HMG1/2与异常结构的DNA的协同结合可能通过溶解结构来去除转录块。在E. coil系统中，HMG蛋白分子中的两个DNA结合域过表达。纯化结构域的三级结构正在研究中。(2) HNG1和HMG2基因的表达与细胞周期和生长的关系：分析HMG1/2基因在细胞周期和生长过程中的表达水平。结果表明，与HMG1基因相比，HMG2基因在细胞周期的G2期表达增强，并依赖于细胞增殖。细胞核中HMG蛋白的相对数量与相关基因的表达水平相关。(3) HMG1蛋白刺激体内转录：研究了HMG1基因在共转染报告基因的细胞中表达的影响。结果表明，HMG1蛋白可刺激共转染基因的转录活性。体内系统可用于有效生产功能活性蛋白。

项目成果

Shigemi Aizawa et al: "DNA-binding protein HNG1 stimulates in vivo transcription" J.Biol. Chem.

Shigemi Aizawa 等人：“DNA 结合蛋白 HNG1 刺激体内转录”J.Biol。

Akiko Yamamoto: "Requirement of two DNA binding domains in HMG1 and 2 for the preferential binding to the supercoiled DNA" Nucleic Acids Res.

Akiko Yamamoto：“HMG1 和 2 中需要两个 DNA 结合域才能优先结合超螺旋 DNA”《核酸研究》。

Akiko Yamamoto,Shou Waga,Hitoshi Shirakawa and Michiteru Yoshida: "Preferential binding of HMG1 protein to the supercoiled DNA." Biochemistry.

Akiko Yamamoto、Shou Waga、Hitoshi Shirakawa 和 Michiteru Yoshida：“HMG1 蛋白优先结合超螺旋 DNA。”

A.Yamamoto et al: "Requirement of two DNA binding domains in HMG1 and 2 for the preferential binding to the supercoiled DNA" Nucleic Acids Res.

A.Yamamoto 等人：“HMG1 和 2 中需要两个 DNA 结合域才能优先结合超螺旋 DNA”Nucleic Acids Res。

Shigemi Aizawa,Hiroaki Nishino,Hitoshi Shirakawa and Michiteru Yoshida: "Stimulation of transient expression of transfected genes by coーtransfection of HMG1 protein cDNA." Molecular and cellular Biology.

Shigemi Aizawa、Hiroaki Nishino、Hitoshi Shirakawa 和 Michiteru Yoshida：“通过共转染 HMG1 蛋白 cDNA 刺激转染基因的瞬时表达”。