Protein phosphatases and cleavage of nucleolin in apoptosis-induced osteoblastic cells

凋亡诱导的成骨细胞中的蛋白磷酸酶和核仁素裂解

• 批准号: 15591936
• 负责人: HANEJI Tatsuji
• 金额: $ 2.24万
• 依托单位: The University of Tokushima
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591936/
• 关键词: apoptosis; osteoblasts; protein dephosphorylation; nucleolin; okadaic acid; nuclear protein; RNAi; 蛋白質脱リン酸化酵素

Protein phosphorylation and dephosphorylation has been recognized as a key mechanism in cell function. Okadaic acid is a potent inhibitor of PP1 and PP2A and induces apoptosis in human or mouse cells including osteoblasts. Nucleolin is an abundantly expressed nucleolar phosphoprotein and is located mainly in the nucleolus. The staining pattern of nucleolin in cultured osteoblastic cells is similar to that of PP1δ. Nucleolin was demonstrated to bind to PP1δ in nucleolus in human osteoblastic cells by using immunocytochemical and immunoprecipitation methods. AgNORs and nucleolin, visible as dots in the nuclei of the control osteoblastic cells, disappeared from the nuclei of the osteoblastic cells treated with okadaic acid. A major band, 110 kDa, was detected in the proteins obtained from osteoblastic cells. The level of the 110 kDa protein decreased in the apoptotic osteoblastic cells, whereas an additional band, 80 kDa, appeared and the level of this protein increased in the proteins prepared from okadaic acid-induced apoptotic osteoblastic cells. The present results indicate that PP1δ directly binds to nucleolin in the nucleolus of osteoblastic cells and that nucleolin is cleaved during apoptosis in osteoblastic cells.

蛋白质磷酸化和去磷酸化已被认为是细胞功能的关键机制。冈田酸是PP1和PP2A的有效抑制剂，可诱导人或小鼠细胞（包括成骨细胞）凋亡。核仁蛋白是一种大量表达的核仁磷酸化蛋白，主要位于核仁中。培养成骨细胞核仁蛋白的染色模式与PP1δ相似。通过免疫细胞化学和免疫沉淀方法证实了人成骨细胞核仁中核蛋白与PP1δ的结合。在对照成骨细胞细胞核中可见的AgNORs和核蛋白在经冈田酸处理的成骨细胞细胞核中消失。在成骨细胞获得的蛋白质中检测到一个主要的条带，110 kDa。在冈田酸诱导的成骨细胞凋亡中，110 kDa蛋白的表达水平降低，而在冈田酸诱导的成骨细胞凋亡中，则出现了一个额外的80 kDa蛋白带，并且该蛋白的表达水平升高。结果表明，PP1δ直接与成骨细胞核仁中的核仁蛋白结合，核仁蛋白在成骨细胞凋亡过程中发生断裂。

项目成果

羽地達次: "骨芽細胞における蛋白質脱リン酸化酵素とニュークレオリン-アポトーシス細胞におけるニュークレオリンの分解-"解剖学会雑誌. 78. 71-75 (2003)

Tatsuji Hanechi：“成骨细胞中的蛋白质去磷酸化酶和核仁素 - 凋亡细胞中核仁素的降解”《解剖学会杂志》78. 71-75 (2003)。

Type 1 protein phosphatase controls biological processes in cultured human osteoblastic cells.

1 型蛋白磷酸酶控制培养的人成骨细胞中的生物过程。

• 发表时间: 2004
• 期刊: Journal of Oral Biosciences 46
• 作者: 小川;郁子 他;Ogawa I. et al.;小川 郁子;Hirohiko Okamura;Hirohiko Okamura;Michi Fujita;Hiroyuki Morimoto
• 通讯作者: Hiroyuki Morimoto

Shinji Kito: "Cleavage of nucleolin and argyrophilic nucleolar organizer region associated proteins in apoptosis-induced cells"Biochemical and Biophysical Research Communications. 300. 950-956 (2003)

Shinji Kito：“凋亡诱导细胞中核仁素和嗜银核仁组织者区域相关蛋白的裂解”生物化学和生物物理研究通讯。

骨芽細胞における蛋白質脱リン酸化酵素とニュークレオリン-アポトーシス細胞におけるニュークレオリンの分解-

成骨细胞中的蛋白质去磷酸化酶和核仁素 - 凋亡细胞中核仁素的降解 -

• 发表时间: 2003
• 期刊: 解剖学雑誌 78
• 作者: 羽地達次

Hiroyuki Morimoto: "Okadaic acid induces apoptosis through double-stranded RNA-dependent protein kinase/eukaryotic initiation factot-2α pathway in human osteoblastic MG63 cells"Journal of Biochemistry. (in press). (2004)

Hiroyuki Morimoto：“冈田酸通过人成骨细胞 MG63 细胞中的双链 RNA 依赖性蛋白激酶/真核起始因子 2α 途径诱导细胞凋亡”《生物化学杂志》（2004 年）。