基础医学与临床 ›› 2017, Vol. 37 ›› Issue (2): 162-168.

• 研究论文 • 上一篇    下一篇

SREBP1c/cm对PERK启动子转录活性及表达的差异性调控分析

胡勤1,毛雨2,陆嘉陵2,谢巍伟3,郑维3,郭风劲4   

  1. 1. 重庆医科大学 基础医学院 细胞生物学及遗传学教研室114信箱
    2. 重庆医科大学基础医学院细胞生物学及遗传学教研室,发育生物学与模式动物平台,重庆
    3. 重庆医科大学 基础医学院 细胞生物学及遗传学教研室
    4. 重庆医科大学
  • 收稿日期:2016-04-18 修回日期:2016-09-26 出版日期:2017-02-05 发布日期:2017-01-16
  • 通讯作者: 郭风劲 E-mail:guofengjin919@vip.sohu.com
  • 基金资助:
    IRE1α与PGRN 双信号通路协同调控小鼠炎性关节炎的发病机制;XBP1S,一个新型的调控软骨细胞分化的转录因子;教育部新世纪优秀人才支持计划(NCET-12-1090)

Differential regulation of SREBP1c/cm on transcriptional activity and expression of PERK promoter

  • Received:2016-04-18 Revised:2016-09-26 Online:2017-02-05 Published:2017-01-16
  • Contact: Fengjin Guo E-mail:guofengjin919@vip.sohu.com

摘要: 目的 探讨转录因子固醇调节元件结合蛋白(SREBP1c)及其活性形式(SREBP1cm)对人蛋白激酶R样内质网激酶(PERK)的调控作用。方法 构建人PERK启动子截短体报告基因载体,与内参质粒pRL-SV40共转入人胚肾细胞HEK293检测荧光素酶活性;用pcDNA3.1(-)-SREBP1c、pcDNA3.1(-)-SREBP1cm与PERK启动子转录活性核心区域共转293T细胞,双荧光素酶报告基因分析SREBP1c及活性形式SREBP1cm对PERK启动子转录活性的调控;RT-PCR和免疫印迹法检测SREBP1c、SREBP1cm对PERK mRNA和蛋白表达的影响。 结果 成功构建PERK启动子截短体报告基因载体,确定了PERK启动子转录活性核心区域;双荧光素酶报告基因分析结果显示SREBP1c抑制PERK启动子的转录活性,而SREBP1cm促进PERK启动子的转录活性。SREBP1c抑制PERK mRNA和蛋白的表达(p<0.05),SREBP1cm促进PERK mRNA和蛋白的表达(p<0.05)。 结论 SREBP1c和SREBP1cm对PERK启动子转录活性及其表达具有相反的调控作用。

关键词: PERK, SREBP1c, 转录活性, ER stress

Abstract: Objective To investigate the effect of sterol regulating element binding protein (SREBP1c) and its active form (SREBP1cm) on human protein kinase R-like endoplasmic reticulum kinase (PERK). Methods Reporter victors of PERK promoter and its truncations are constructed with pGL3-basic and co-transfected with internal reference pRL-SV40 into cell and luciferase activity was detected. pcDNA3.1(-)-SREBP1c or pcDNA3.1(-)-SREBP1cm was co-transfected with PERK promoter transcriptional activity core regions and the detection of dual-luciferase reporter gene was used to analyze the regulation of SREBP1c/1cm on PERK promoter transcriptional activity. The expression level of PERK mRNA and protein were detected by RT-PCR and Western blot. Results PERK promoter and truncations were successfully constructed into pGL3-basic, and PERK promoter core area of transcriptional activity had determined; Dual-luciferase report gene showed that SREBP1c inhibits PERK promoter transcriptional activity and SREBP1cm promotes PERK promoter transcriptional activity. RT-PCR and Western blot showed that SREBP1c decreased PERK mRNA and protein expression, but SREBP1cm increased PERK mRNA and protein expression, which was consistent with the detection of dual-luciferase report gene. Conclusions SREBP1c and SREBP1cm have a opposite regulation effect on PERK promoter transcriptional activity and its expression.

Key words: PERK, SREBP1c, Transcriptional activity, ER stress