摘要
目的明确转录因子Sp1在人内皮细胞高表达脂多糖相关因子1(endothelial-overexpressed lipopolysiccha-ride-associated factor 1,EOLA1)基因转录调控中的作用。方法定点突变EOLA1基因核心启动子-734～-666序列中推断的Sp1结合位点,观察启动子活性变化;进行凝胶电泳迁移和染色质免疫共沉淀验证Sp1转录因子与启动子区顺式作用元件发生相互作用。结果突变Sp1结合位点后,核心启动子的活性(0.12±0.01)较野生启动子(2.51±0.98)显著下降(P<0.05),EMSA和染色质免疫共沉淀验证了SP1与启动子区顺式作用元件相互作用。结论转录因子Sp1调控了EOLA1基因的基础转录。
Objective To investigate the role of stimulating protein 1（Sp1） transcription factor in the transcriptional regulation of human endothelial-overexpressed lipopolysaccharide-associated factor 1（EOLA1） gene.Methods Site-mutation of a putative Sp1 regulatory site locating at-734 to-666 of the core promoter of EOLA1 gene was used to analyze promoter activity.Electrophoretic mobility shift assay（EMSA） and chromatin immunoprecipitation analysis（ChIP） were performed to confirm whether Sp1 bound to the site.Results Mutation of the Sp1 regulatory site resulted in a significant decline of EOLA1 promoter activity（P〈0.05）.EMSA and ChIP showed that Sp1 bound to the site.Conclusion Basal transcription of EOLA1 gene is regulated by Sp1 transcription factor.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2011年第21期2249-2251,共3页
Journal of Third Military Medical University
基金
国家自然科学基金(30670838)~~
关键词
EOLA1
SP1
启动子
凝胶电泳迁移
染色质共沉淀
endothelial-overexpressed lipopolysaccharide-associated factor 1
stimulating protein 1
promoter
electrophoretic mobility shift assay
chromatin immunoprecipitation analysis
