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Functional analysis of ubiquitin ligase by knock-out mise

通过敲除mise对泛素连接酶进行功能分析

基本信息

批准号：
12480211
负责人：
NAKAYAMA Keiko
金额：
$ 9.34万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12480211/
关键词：
Ubiquitin Ligase knock-out mouse Skp2 Cul-1 Skp1 p27 F-box protein SCF複合体中心体ユビキチン化

项目摘要

1) We have reported that F-box protein, FWD1 works as a ubiquitin ligase on the degradation of lκB or β-catenin. FWD1 constitutes SCF complex with Cul-1, Skp1, and Rbx1. In this study, we generated knock-out mice of Cul-1 and Skp1, which are components of SCF complex, and another F-box protein, Skp2. Analysis of these mice shows the biological role of protein degradation by SCF complex system, one category of proteolysis by ubiquitin-proteasome system.2) Skp2 is a ubiquitin ligase of cyclin E and p27^<Kip1>. In Skp2 knock-out mice, cyclin E and p27^<Kip1> was accumulated abnormally. Cells in the mutant mice contain markedly enlarged nuclei with polyploidy and multiple centrosomes, and show a reduced growth rate and increased apoptosis.3) We generated and characterized mice lacking both Skp2 and p27^<Kip1>. The Skp2^<-/->p27^<-/-> mice did not exhibit the overreplication phenotype, suggesting hat p27^<Kip1> accumulation is required for its development.4) Cul-1 and Skp1 knock-out mice are died between on embryonic day 5.5 and 6.5. In these embryos, cyclin E is accumulated. It is postulated that Cul-1 and Skp1 are common components of SCF complex and relate to degradation of many kinds of protein. Early embryonic death of knock-out mice supports that SCF complex participate in many kinds of protein degradation system.

1)我们已经报道了F-盒蛋白FWD_1在LκB或β-连环蛋白的降解中起泛素连接酶的作用。FWD1与Cul-1、Skp1、Rbx1构成SCF复合体。在这项研究中，我们产生了SCF复合体的组成成分Cul-1和Skp1以及另一种F-box蛋白Skp2的敲除小鼠。对这些小鼠的分析显示了SCF复合系统对蛋白质降解的生物学作用，这是泛素-蛋白酶体系统的一种蛋白质分解。2)Skp2是细胞周期蛋白E和p27^&lt；Kip1&gt；的泛素连接酶。在Skp2基因敲除小鼠中，细胞周期蛋白E和p27^&lt；Kip1；突变小鼠的细胞核明显增大，具有多倍体和多个中心体，生长速度减慢，细胞凋亡率增加。3)我们建立并鉴定了同时缺失Skp2和p27^&lt；Kip1&gt；Skp2^&lt；-/-&gt；p27^&gt；-/-&gt；小鼠没有表现出过度复制的表型，这表明它的发育需要p27^&lt；Kip1&gt；积累。4)Cul-1和Skp1基因敲除小鼠在胚胎5.5天到6.5天之间死亡。在这些胚胎中，细胞周期蛋白E积累。推测Cul-1和Skp1是SCF复合体的共同成分，与多种蛋白质的降解有关。基因敲除小鼠早期胚胎死亡支持SCF复合体参与多种蛋白质降解系统。