血小板源生长因子B链基因上游序列HMGI结合区的初步鉴定  

Characterization of the HMGI-binding AT-rich regions in the upstream sequence of the platelet-derived growth factor-B chain gene

作　　者：王小明[1] 邱劲[1] 石建茹[1] 斯琴[1] 李素敏[1] 沈传陆[1] 郭恒怡[1] 吴其夏[1] 

机构地区：[1]中国医学科学院基础医学研究所中国协和医科大学基础医学院病理生理系,北京100005

出　　处：《中国病理生理杂志》2006年第2期257-261,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金青年基金资助项目(No.39800062);国家自然科学基金重点项目(No.39730220)

摘　　要：目的:观察转录结构因子高迁移率蛋白I(HMGI)与血小板源生长因子B链(PDGF-B)基因上游序列的结合并初步鉴定HMGI的结合序列。方法:应用凝胶迁移率改变法(EMSA)观察重组人HMGI蛋白与PDGF-B基因上游-1758/+43bp片段的结合,并逐步确定与其结合的AT富含序列。结果:重组人HMGI蛋白能较特异地与PDGF-B链基因上游-1758/+43bp片段结合,分离到的两个HMGI结合片段,-1393/-1181bp和-190/+43bp,均含有AT富含序列TTTATAAA(-1333/-1326bp,-1314/-1307bp和-30/23bp)。HMGI能与含TT-TATAAA的合成寡核苷酸结合,并能促进转录因子NF-κB与拼接于旁侧的PDGF-B基因的切应力反应元件结合。结论:HMGI在体外能与PDGF-B链基因上游的TTTATAAA序列结合,可能参与PDGF-B基因的转录调控。AIM： To determine whether the high mobility group protein I （HMGI） is able to bind to the upstream sequence of platelet - derived growth factor B - chain gene and to characterize the HMGI - binding AT- rich regions. METHODS： Recombinant human HMGI （rhHMGI） protein was prepared and electrophoresis mobility shift assay （EMSA） was used, RE- SULTS： The binding of rhHMGI to PDGF - B （ - 1 758/＋ 43 bp） was observed in vitro. Two major HMGI - binding fragments - 1 392/- 1 180 bp and - 188/＋ 43 bp were identified, which contained the same AT - rich sequence TTTATAAA （ - 1 333/ - 1 326 bp, - 1 314/- 1 307 bp and - 30/- 23 bp）. An oligonucleotide bound to the TITATAAA and the GAGACC, the core sequence of the shear stress response element of the PDGF- B, could also bind to the HMGI. Furthermore, HMGI facilitated the binding of NF - κB to the GAGACC in the oligonucleotide. CONCLUSION： The HMGI could bind to the upstream sequence of the PDGF- B gene via the AT- rich sequence TITATAAA, which may play a role in the transcriptional regulation of the PDGF- B gene.

关 键 词：高迁移率族蛋白质类 基因 血小板源性生长因子-B链 

分 类 号：R363[医药卫生—病理学]