Gene Expression of Regulatory Protein Regucalcin of Ca^<2+>-Signaling and Its Regulatory Mechanism.

Ca^2-信号调节蛋白Reguccalcin的基因表达及其调节机制。

• 批准号: 08672522
• 负责人: YAMAGUCHI Masayoshi
• 金额: $ 1.47万
• 依托单位: University of Shizuoka
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08672522/
• 关键词: Regucalcin; Regulation of gene expression; Liver nuclear; Calcium signaling; Transcription regulation; レギュカルチン遺伝子; カルシウム; 遺伝子発現

This investigation was undertaken to clarify the gene expression and its regulatory mechanism of regucalcin which can regulate Ca^<2+> signalin in liver cells due to hormonal stimulation. The following results were obtained.1.Regucalcin could inhibit RNA synthesis in the nuclei of normal rat liver and regenerating rat liver.2.Regucalcin mRNA was expressed in the transformed HepG2 cells and its expression was stimulated by insulin.3.The cDNA clone encoding a regucalcin was isolated from a mouse liver cDNA library and sequenced. Regucalcin mRNA in mice was uniquely expressed in the liver, and that its expression was stimulated by calcium administration.4.The existence of nuclear factors which bind to the 5'-flanking region of regucalcin gene in rats was found. These identified components was involved in the tissue-specific regulation of regucalcin gene expression.5.When nuclear proteins obtained from various rat tissues were used in gel mobility-shift assays, tissue-specific formation of a protein-DNA complex was found in the liver and kidney. Ca^<2+>-induced binding of the AP-1 factor to the regucalcin gene was completely inhibited by simultaneous administration of an antagonist of calmodulin. The 5'-flanking region of the rat regucalcin gene ligated to a luciferase reporter gene possessed the promotor activity in H4-II-E hepatoma cells.

本研究旨在阐明regcalcin的基因表达及其调控机制，regcalcin可以调节肝细胞中由于激素刺激而产生的Ca 2+ 信号素。得到以下结果：1.Reguccalcin能够抑制正常大鼠肝脏和再生大鼠肝脏细胞核内的RNA合成。2.Reguccalcin mRNA在转化的HepG2细胞中表达，并且其表达受胰岛素刺激。3.从小鼠肝脏cDNA文库中分离编码regcalcin的cDNA克隆并测序。小鼠Reguccalcin mRNA在肝脏中独特表达，钙剂可刺激其表达。4.大鼠体内存在与Reguccalcin基因5'侧翼区结合的核因子。这些鉴定出的成分参与了regcalcin基因表达的组织特异性调节。5.当从不同大鼠组织获得的核蛋白用于凝胶迁移率变化测定时，在肝脏和肾脏中发现了组织特异性形成的蛋白质-DNA复合物。 Ca 2+ 诱导的AP-1因子与调节钙蛋白基因的结合被同时施用钙调蛋白拮抗剂完全抑制。与荧光素酶报告基因连接的大鼠调节钙素基因的 5'-侧翼区域在 H4-II-E 肝癌细胞中具有启动子活性。

项目成果

Murata,Tomiyasu: "Ca^<2+> administration stimulates the binding of AP-1 factor to the S^1-flanking region of the rat gene for the Ca^<2+>-binding protein regucalor" Biochemical Journal. 329(1). 157-163 (1998)

Murata，Tomiyasu：“Ca ^ 2 施用刺激AP-1因子与Ca ^ 2 结合蛋白调节剂的大鼠基因的S ^ 1侧翼区域的结合”生物化学杂志。

Murata, Tomiyasu: "Ca^<2+> administration stimulates the binding of AP-1 factor to the S'-flanking region of the rat gene for the Ca^<2+>-binding protein regular." Biochemical Journal. 329. 157-163 (1998)

Murata, Tomiyasu：“Ca^2 给药刺激 AP-1 因子与大鼠基因的 S 侧翼区域结合，形成 Ca^2 结合蛋白常规。”

Tomiyasu Murata, Nobuko Synya, Masayoshi Yamaguchi: "Expression of calcium-binding protein regucalcin mRNA in the cloned human hepatoma cells (HepG2) : Stimulation by insulin." Molecular and Cellular Biochemistry. 175. 163-168 (1997)

Tomiyasu Murata、Nobuko Synya、Masayoshi Yamaguchi：“克隆人肝癌细胞 (HepG2) 中钙结合蛋白 regucalcin mRNA 的表达：胰岛素刺激。”

Tomiyasu Murata, Masayoshi Yamaguchi: "Tissue-specific binding of nuclear factors to the 5'-flanking region of rat gene for calcium-binding protein regucalcin." Molecular and Cellular Biochemistry. (in press). (1998)

Tomiyasu Murata、Masayoshi Yamaguchi：“核因子与钙结合蛋白调节钙蛋白大鼠基因 5 侧翼区域的组织特异性结合。”

Murata, Tomiyasu: "Molecular cloning of the cDNA coding for reigucalcir and its mRNA expression in mouse liver : the expression is stimulated by colium" Molecular and Cellular Biochemistry. (印刷中). (1997)

Murata, Tomiyasu：“小鼠肝脏中编码瑞古卡西尔的 cDNA 的分子克隆及其 mRNA 表达：结肠刺激表达”《分子与细胞生物化学》（出版中）。