Mdecular cloning and analysis of S' -flanking region of human MYPT1 gene

人MYPT1基因S侧翼区的分子克隆与分析

• 批准号: 10670645
• 负责人: IRO Masaaki
• 金额: $ 2.05万
• 依托单位: Mie University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670645/
• 关键词: Myosin Phosphatase; MYPT1; Promoter; Sp1; Human Gene; 転写活性調節領域

MYPT1, a subunit of myosin phosphatase, has critical functions including activation and regulation of phosphatase activity.We have cloned and sequenced a 5 kb genomic fragment in the 5'-flanking region of the human MYPT1 gene. The transcription initiation site (+1) was 268 bp upstream from the translation start site. In this promoter there are no canonical TATA or CAAT box elements but there is a high GC-rich sequence (-58 to -64). Basal promoter activity was due to the GC-rich region that contained one Sp 1 transcription factor-binding site. The specific interaction of Sp 1 with this region was demonstrated by the electrophoresis mobility shift assay. Therefore, it was concluded that the MYPT1 gene is a housekeeping gene. Luciferase reporter assays showed the presence of two regions for positive elements (-246 to -230 and -170 to -198) and one for a negative element (-199 to -229).

MYPT 1是肌球蛋白磷酸酶的一个亚基，具有激活和调节磷酸酶活性的重要功能，我们克隆了人MYPT 1基因5 '端侧翼区一个5 kb的基因组片段，并进行了序列测定。转录起始位点（+1）位于翻译起始位点上游268 bp处。在该启动子中，不存在典型的TATA或CAAT盒元件，但存在高GC富集序列（-58至-64）。基础启动子活性是由于富含GC的区域含有一个Sp 1转录因子结合位点。电泳迁移率变动分析证实了Sp1与该区域的特异性相互作用。因此，可以得出结论，MYPT 1基因是管家基因。荧光素酶报告基因测定显示存在两个阳性元件区域（-246至-230和-170至-198）和一个阴性元件区域（-199至-229）。

项目成果

Jianhua Feng: "Dephesphorylation of distinct sites on the 20KDa myosin light chain by smooth muscle myosin phosphatase"FEBS Letters. 448. 101-104 (1999)

冯建华：“平滑肌肌球蛋白磷酸酶对 20KDa 肌球蛋白轻链上不同位点的去磷酸化”FEBS Letters。

Jianhua Feng: "Inhibitory phosphorylation site for Rho-associated kinase on smooth muscle phosphatase"Journal of Biological Chemistry. 274. 37385-27390 (1999)

冯建华：“平滑肌磷酸酶上 Rho 相关激酶的抑制性磷酸化位点”生物化学杂志。

Hirokazu Kotani: "The δ isoform of protein phosphatase type 1 is localized in nucleolus and dephosphory lates nucleolar proteins" Biochemical and Biophysical Research Communications. 249. 292-296 (1998)

Hirokazu Kotani：“1 型蛋白磷酸酶的 δ 同工型位于核仁和去磷酸化核仁蛋白”《生物化学和生物物理研究通讯》249. 292-296 (1998)。

Jianhua Feng: "Inhibitory phosphorylation site for Rho-associated kinase on smooth muscle myosin phosphatese"Journal of Biological Chemistry. 274. 37385-37390 (1999)

冯建华：“平滑肌肌球蛋白磷酸酶上 Rho 相关激酶的抑制性磷酸化位点”生物化学杂志。

Howard K.Surks: "Specific association of CGMP-dependent protein kinase with the myosin binding subunit of myosin phosphatase"Science. 286. 1583-1587 (1999)

Howard K.Surks：“CGMP 依赖性蛋白激酶与肌球蛋白磷酸酶的肌球蛋白结合亚基的特异性关联”科学。