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Function of DADl Protein in Programd Cell Death

DAD1蛋白在程序性细胞死亡中的功能

基本信息

批准号：
07044203
负责人：
NISHIMOTO Takeharu
金额：
$ 1.28万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1995
资助国家：
日本
起止时间：
1995 至无数据
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07044203/
关键词：
Cell death Ts Mutation DAD1 Glycosylation Modification of protein OST2 C.elegance

项目摘要

We have isolated a series of temperature sensitive growth mutant from the hamster BHK21 cell line. Among those ts mutants, we found the mutants which enter apoptotic cell death at the restrictive temperature. In one of them tsBN7, chromosomal DNA was fragmented with a typical manner like apoptptic cell death depending on protein synthesis. We have cloned a human gene complementing tsBN7 mutation and designated as DAD1 encoding a protein of 12.5 kDa which is well conserved through evolution. As a co-work, we have cloned a dadl homologue of Caenorhabditis elegans (Ce-dadl). Ce-dadl is more than 60% identical to vertebrate DAD1 in amino acid level and well complemented tsBN7 mutation. By overexpression, human DAD1 suppressed programd cell death in C.elegance.Recently, DAD1 was found to be a homologue of yeast OST2 involved in N linked glycosylation. Based on this finding, we found that a glycosylation of p450 is temperature sensitive in tsBN7 cells. As reported that N linked glycosylation occurs prior to entering Golgi apparatus, DAD1 seems to be located in endoplasmic reticulum. We don't know how a defect of N linked glycosylation causes apoptotic cell death. In order to clarify this issue, we are deleting DAD1 gene from mouse chromosome by gene targeting.

我们从仓鼠BHK21细胞系中分离出一系列温度敏感生长突变体。在这些突变体中，我们发现了在限制温度下进入凋亡细胞死亡的突变体。在其中一个tsBN7中，染色体DNA以一种典型的方式断裂，就像依赖于蛋白质合成的凋亡细胞死亡一样。我们克隆了一个与tsBN7突变互补的人类基因，并将其命名为DAD1，编码一个12.5 kDa的蛋白，该蛋白在进化过程中具有良好的保守性。作为合作研究，我们克隆了秀丽隐杆线虫（cenorhabditis elegans, Ce-dadl）的dadl同源物。在氨基酸水平上，ce - DAD1与脊椎动物DAD1有60%以上的相同，并且很好地补充了tsBN7突变。通过过表达，人类DAD1抑制了C.elegance的程序性细胞死亡。最近，DAD1被发现是酵母OST2参与N连锁糖基化的同源物。基于这一发现，我们发现p450的糖基化在tsBN7细胞中对温度敏感。据报道，N链糖基化发生在进入高尔基体之前，DAD1似乎位于内质网。我们不知道N链糖基化缺陷是如何导致细胞凋亡的。为了澄清这一问题，我们正在通过基因靶向从小鼠染色体上删除DAD1基因。