[1]刘小平,巩丽云,王亮,等.ATF1与CDK3蛋白相互作用研究[J].深圳大学学报理工版,2010,27(4):497-501.
 LIU Xiao-ping,GONG Li-yun,WANG Liang,et al.Interaction analysis of ATF1 and CDK3[J].Journal of Shenzhen University Science and Engineering,2010,27(4):497-501.
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ATF1与CDK3蛋白相互作用研究()
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《深圳大学学报理工版》[ISSN:1000-2618/CN:44-1401/N]

卷:
第27卷
期数:
2010年4期
页码:
497-501
栏目:
生命科学
出版日期:
2010-10-31

文章信息/Info

Title:
Interaction analysis of ATF1 and CDK3
文章编号:
1000-2618(2010)04-0497-05
作者:
刘小平1巩丽云2王亮2谭志平1郑多2
1)中南大学湘雅二医院,长沙 410011
2)深圳大学医学院,深圳 518060
Author(s):
LIU Xiao-ping1GONG Li-yun2WANG Liang2TAN Zhi-ping1and ZHENG Duo2
1)Xiangya Second Hospital, Central South University, Changsha 410011, P.R.China
2)College of Medicine, Shenzhen University, Shenzhen 518060, P.R.China
关键词:
细胞转化生物大分子结构与功能转录激活因子1细胞周期蛋白依赖性蛋白激酶3真核细胞双杂交系统HEK293细胞
Keywords:
cell transformationstructure and function of biomacromoleculesactivating transcription factor 1(ATF1)cyclin-dependent kinase 3(CDK3)mammalian two-hybrid assayHEK293 cells
分类号:
Q 291;Q 71
文献标志码:
A
摘要:
研究真核细胞转录激活因子1(activating transcription factor 1,ATF1)与细胞周期蛋白依赖性蛋白激酶3(cyclin-dependent kinase 3,CDK3)的相互作用机制,分别构建ATF1的激酶诱导区域(kinase inducible domain,KID)结构域、谷氨酰胺富含域(glutamine rich domain,Q2)结构域及亮氨酸拉链区(basic leucine zipper domain,bZIP)结构域缺失体与VP16的融合表达载体,在HEK293细胞过表达,应用真核细胞双杂交系统,研究ATF1结构域与CDK3的相互作用.结果表明,ATF1全蛋白与CDK3蛋白之间存在强烈的相互作用;ATF1结构域缺失体与VP16的融合蛋白在HEK293细胞中可获得高表达,但ATF1的任何一种结构缺失体与CDK3蛋白结合都会严重减弱其与CDK3蛋白的相互作用,只有完整的ATF1蛋白才能实现与上游CDK3蛋白的结合.
Abstract:
To study the interaction mechanism between activating transcription factor 1(ATF1)and cyclin-dependent kinase 3 (CDK3),the plasmids for expression of VP16-tagged ATF1 and the VP16-ATF1 deletions(residues 1~57,1~214,or 215~271) were constructed.HEK293 cells were transfected and proteins overexpressed were detected by Western blotting.Mammalian two-hybrid assay was conducted and luciferase activity was measured.Results show that an 11-fold increase in interacting activity between pACT-ATF1 and pBIND-CDK3 compared with the basal level (value of 1) of pG5-luc/pBIND-CDK3/pACT control.Neither of ATF1 deletion mutants show strong interaction with CDK3,suggesting that the full length of ATF1 is required for the interaction with CDK3.

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备注/Memo

备注/Memo:
收稿日期:2010-06-22;修回日期:2010-09-10
基金项目:国家自然科学基金资助项目(30871247),深圳市科技计划基金资助项目(Jc200903120125)
作者简介:刘小平(1967-),男(汉族),湖南省汨罗市人,中南大学湘雅二医院主治医师.E-mail:liupx67@hotmail.com
通讯作者:郑多(1968-),女(汉族),深圳大学教授、博士.E-mail:dzheng@szu.edu.cn
更新日期/Last Update: 2010-11-09