ULK1启动子区含有抑制ULK1转录的G4结构  

ULK1 promoter region contains G-quadruplexes that inhibit transcrip⁃tion of ULK1

作　　者：刘雪萍 黄伟伟[1] 李晓梅 LIU Xue-ping;HUANG Wei-wei;LI Xiao-mei(College of Life Science,Northwest Agriculture&Forestry University,Yangling 712100,China)

机构地区：[1]西北农林科技大学生命科学学院,陕西杨凌712100

出　　处：《中国病理生理杂志》2021年第7期1270-1276,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.31300654)。

摘　　要：目的:研究G四联体(G-quadruplex,G4)结构对UNC-51样激酶1(UNC-51-like kinase 1,ULK1)基因的转录调控作用。方法:分析ULK1转录起始点上游1000 bp启动子序列的G/C含量;预测ULK1启动子区G4形成序列,利用圆二色谱、非变性聚丙烯酰胺凝胶电泳和DNA银染鉴定G4;通过构建ULK1启动子报告基因质粒和用G4配体处理人前列腺癌DU-145细胞,结合报告基因检测、RT-qPCR和Western blot实验检测G4对ULK1转录的调控作用。结果:ULK1启动子区G/C含量丰富;预测发现10条G4形成序列;体外实验表明G4形成序列可以形成G4结构;细胞实验表明ULK1启动子区的G4结构负调控ULK1的转录。结论:ULK1启动子区可形成抑制ULK1转录的G4结构。这为揭示ULK1的转录调控机制提供了新的线索。AIM:To study the regulatory effect of G-quadruplex(G4)structure on the transcription of UNC-51-like kinase 1(ULK1)gene.METHODS:The G/C content in ULK1 promoter sequence,1000 bp upstream of the transcription start point,was analyzed.The G4 sequence in the ULK1 promoter region was predicted,and the G4 structure was identified by circular dichroism,non-denaturing polyacrylamide gel electrophoresis and DNA silver staining.After the ULK1 promoter reporter plasmid was constructed and human prostate carcinoma DU-145 cells were treated with G4 ligand,reporter gene detection,RT-qPCR and Western blot experiments were used to identify the regulatory effect of G4 on ULK1 transcription.RESULTS:The G/C content of ULK1 promoter region was abundant,and ten G4-forming sequences were predicted to be found.In vitro experiments showed that G4-forming sequences formed G4 structures.The cell experiments showed that the G4 structure of ULK1 promoter region negatively regulated ULK1 transcription.CONCLUSION:The pro-moter region of ULK1 forms G4 structures that inhibit ULK1 transcription,which provides new clues for revealing the tran-scriptional regulatory mechanism of ULK1.

关 键 词：UNC-51样激酶1 G四联体 圆二色谱 转录调控 

分 类 号：R363[医药卫生—病理学] R329.2[医药卫生—基础医学]