转录因子COUP-TFII对端粒酶活性的抑制及其相互作用蛋白的分离和鉴定  

The transcription factor COUP-TFII inhibits telomerase activity and the separation and identification of its interactional proteins

作　　者：王强[1] 白增亮[1] 李璇[2] 侯琳[2] 张波[2] 

机构地区：[1]山东大学生命科学学院,山东济南250100 [2]北京大学医学部病理系,北京100083

出　　处：《山东大学学报（理学版）》2004年第4期95-100,共6页Journal of Shandong University(Natural Science)

基　　金：国家自然科学基金资助项目 ( 3 0 170 3 64 );北京市 2 48工程生物领域资助项目 (H0 10 2 10 47112 );北京大学人类疾病基因研究中心资助项目 ( 2 0 0 2 111)

摘　　要：端粒酶催化亚基hTERT的转录调控是端粒酶活性调节的关键步骤 .实验分离和克隆了参与hTERT转录调控的转录因子COUP TFII,分析了其对HeLa细胞端粒酶活性的生物学效应 ,并分离和鉴定了与其相互作用的蛋白质 .结果表明在以hTERT启动子为诱饵的酵母单杂交体系中 ,COUP TFII可以与hTERT启动子结合 ;得到了纯度高达98%的COUP TFII融合蛋白 ,并且其结合于hTERT启动子 2 0 1～ +35区段 ,抑制HeLa细胞端粒酶的活性 ;从HeLa细胞核蛋白中分离到了 2种与COUP TFII相互作用的蛋白质 .研究初步揭示了COUP TFII在细胞分化和肿瘤转化过程中的参与途径和调控机制 ,为开启以端粒酶为靶目标的肿瘤治疗奠定了基础 .hTERT transcription is the key step in the regulation of telomerase activity. The transcription factor COUP-TFII which involved in regulating the transcription of hTERT was identified and cloned, and its role in telomerase activity in HeLa cells was studied. Its interactional proteins were also separated from HeLa cell nuclear extracts in the present research. COUP-TFII was proved to interact with hTERT promoter in hTERT promoter-based yeast one-hybrid assay. COUP-TFII fusion protein was purified to the fineness ratio up to 98% and it could firmly bind to -201 to +35 fragment of hTERT promoter, and inhibit telomerase activity; two proteins which interact with COUP-TFII were separated from HeLa cell nuclear extracts. The results provide a new clue to profile of COUP-TFII in cellular differentiation and cancer transformation,and will be useful to open up a new strategy for telomerase-targeted anticancer therapy.

关 键 词：酵母单杂交 COUP-TFII 亲和层析 端粒酶 相互作用蛋白 

分 类 号：Q279[生物学—细胞生物学]