[1]冯波,曾虹燕,钟英丽,等.转录因子AP-2α与TES蛋白相互作用分析[J].深圳大学学报理工版,2008,25(4):432-436.
 FENG Bo,ZENG Hong-yan,ZHONG Ying-li,et al.Analysis of interaction of AP-2α and TES[J].Journal of Shenzhen University Science and Engineering,2008,25(4):432-436.
点击复制

转录因子AP-2α与TES蛋白相互作用分析()
分享到:

《深圳大学学报理工版》[ISSN:1000-2618/CN:44-1401/N]

卷:
第25卷
期数:
2008年4期
页码:
432-436
栏目:
生命科学
出版日期:
2008-10-31

文章信息/Info

Title:
Analysis of interaction of AP-2α and TES
文章编号:
1000-2618(2008)04-0432-05
作者:
冯波1曾虹燕1钟英丽2张健2
1)湘潭大学生物工程系,湘潭 411105
2)湖南师范大学生命科学学院,长沙 410081
Author(s):
FENG Bo1ZENG Hong-yan1ZHONG Ying-li2and ZHANG Jian2
1)Department of Biotechnology,Xiangtan University,Xiangtan 411105,P.R.China
2)College of Life Science,Hunan Normal University,Changsha 410081,P.R.China
关键词:
GST Pull-down酵母双杂交AP-2αTESLIM结构域
Keywords:
GST pull-downyeast two-hybridAP-2αTESLIM domain
分类号:
Q 58;Q 503
文献标志码:
A
摘要:
为研究转录因子AP-2α与TES蛋白的相互作用机制,分别构建AP-2α与GST的融合蛋白原核表达载体及TES与His的融合蛋白原核表达载体,在大肠杆菌E.coli BL 21中诱导表达,表达产物经纯化后用GST Pull-down及免疫印迹技术分析其相互作用关系.实验结果表明,AP-2α与TES之间存在相互作用是由于AP-2α C端能与TES C端的首个LIM结构域结合,酵母双杂交实验也证实了该结果;同时根据AP-2α只能与TES C端而不能与TES全长蛋白结合的实验事实提出了TES蛋白存在自身相互作用的可能,这种自身折叠封闭了位于TES C端首个LIM结构域上的AP-2α结合位点.
Abstract:
To study the interaction mechanism between transcription factor AP-2α and TES protein,prokaryotic vectors for expressing GST-AP-2α and His-TES were constructed respectively.Fusion proteins were induced in E.coli BL 21 and then were analyzed with GST pull-down assay.Results show that the interaction between TES and AP-2α is due to AP-2α C-terminus binding with the first LIM domain of TES C-terminus.These results were also confirmed in yeast two-hybrid system.Furthermore,a possible self-interaction of TES protein,which could block the binding site for AP-2α in the first LIM domain of TES,is suggested,based on the fact that AP-2α interacted with TES C-terminus instead of TES full-length protein.

参考文献/References:

[1]Williams T,Tjian R.一个AP-2二聚体模序的结构特征及其异性DNA结合蛋白功能[J].科学,1991,251(4997):1067-1071(英文版).
[2]Ruiz M,Pettaway C,Song R,等.前列腺癌细胞中AP-2α的抑癌性并阻止血管内皮生长因子的转录[J].癌症研究,2004,64(2):631-638(英文版).
[3]张健,Donaldson S,Serbedzija G,等.转录因子AP-2缺失鼠的神经管,骨骼和体壁缺陷 [J].自然,1996,381(6579):238-241(英文版).
[4]张健,Brewer S,Huang J,等.转录因子AP-2α过表达抑制乳腺的生长与形成[J].发育生物学,2003,256(1):128-146(英文版).
[5]钟英丽,刘云海,胡翔,等.用酵母双杂交筛选与AP-2α相互作用的因子[J].湖南师范大学自然科学学报,2003,26(3):73-77.
[6]Tobias E S,Hurlstone A F,MacKenzie E,等.位于7q31.1的TES基因在肿瘤中被甲基化并编码一种新型的生长抑制LIM结构域蛋白 [J].肿瘤基因杂志,2001,20(22):2844-2853(英文版).
[7]Alessandra D,Nicola Z,Claudia R,等.基因敲除鼠显示Testin具有抑癌功能[J].美国科学院院报,2005,102(31):10947-10951(英文版).
[8]Marshak D R,Kadonaga J T,Burgess R R,等.蛋白质纯化与鉴定实验指南[M].朱厚础,译. 北京:科学出版社,2002.
[9]唐玉林.ABA信号元件ABI1和AtGluRS间相互作用分析[J].深圳大学学报理工版,2004,21(4):319-323.
[10]Leena L,Paivi M,Marikki L.细胞支架相互作用LIM结构域蛋白CRP1抑制细胞增殖并保护细胞免遭胁迫诱导性凋亡[J].实验细胞研究,2007,313(24):4236-4242(英文版).
[11]Yuan C,Meizan L,Yuka M H,等.LIM结构域在PKC-ENH-N型Ca2+通道复合体形成过程中的重要作用[J].细胞信号,2006,18(2):215-224(英文版).

[1]Williams T,Tjian R.Characterization of a dimerization motif in AP-2 and its function in heterologous DNA-binding proteins[J].Science,1991,251(4997):1067-1071.
[2]Ruiz M,Pettaway C,Song R,et al.Activator protein 2α inhibits tumorigenicity and represses vascular endothelial growth factor transcription in prostate cancer cells[J].Cancer Research,2004,64(2):631-638.
[3]ZHANG Jian,Donaldson S,Serbedzija G,et al.Neural tube,skeletal and body wall defects in mice lacking transcription factor AP-2[J].Nature,1996,381(6579):238-241.
[4]ZHANG Jian,Brewer S,Huang J,et al.Overexpression of transcription factor AP-2 alpha suppresses mammary gland growth and morphogenesis[J].Developmental Biology,2003,256(1):128-146.
[5]ZHONG Ying-li,LIU Yun-hai,HU Xiang,et al.Identify the interacting partners of AP-2α using yeast two-hybrid system[J].Journal of Natural Science of Hunan Normal University,2003,26(3):73-77 (in Chinese).
[6]Tobias E S,Hurlstone A F,MacKenzie E,et al.The TES gene at 7q31.1 is methylated in tumours and encodes a novel growth-suppressing LIM domain protein[J].Oncogene,2001,20(22):2844-2853.
[7]Alessandra D,Nicola Z,Claudia R,et al.Knockout mice reveal a tumor suppressor function for Testin[J].PNAS,2005,102(31):10947-10951.
[8]Marshak D R,Kadonaga J T,Burgess R R,et al.Strategies for Protein Purification and Characterization:a Laboratory Course Manual[M].ZHU Hou-chu,trans.Beijing:Science Press,2002 (in Chinese).
[9]TANG Yu-lin.Analysis of the interaction of the ABA signaling components ABI1 and AtGluRS[J].Journal of Shenzhen University Science and Engineering,2004,21(4):319-323 (in Chinese).
[10]Leena L,Paivi M,Marikki L.Cytoskeleton-interacting LIM-domain protein CRP1 suppresses cell proliferation and protects from stress-induced cell death [J].Experimental Cell Research,2007,313(24):4236-4242.
[11]Yuan C,Meizan L,Yuka M H,et al.Essential role of the LIM domain in the formation of the PKC-ENH-N-type Ca2+ channel complex [J].Cellular Signalling,2006,18(2):215-224.

相似文献/References:

[1]欧阳秋玲,刘士德,张建华,等.采用酵母双杂交法筛选PSRPK相关蛋白基因[J].深圳大学学报理工版,2006,23(3):222.
 OUYANG Qiu-ling,LIU Shi-de,ZHANG Jian-hua,et al.Screening of the psrpk一related protein genes with yeast two-hybrid method[J].Journal of Shenzhen University Science and Engineering,2006,23(4):222.

备注/Memo

备注/Memo:
收稿日期:2007-12-13;修回日期:2008-08-23
基金项目:国家自然科学基金资助项目(30270723,30470945);湖南省教育厅资助项目(06C816)
作者简介:冯波(1976-),男(汉族),湖南省湘乡市人,湘潭大学讲师.E-mail:fengbo@xtu.edu.cn
通讯作者:张健(1963-),男(汉族),湖南师范大学教授、博士生导师.E-mail:zhangjian@hunnu.edu.cn
更新日期/Last Update: 2008-11-26