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ER stress activation impairs the expression of circadian clock and clock-controlled genes in NIH3T3 cells via an ATF4-dependent mechanism

机译:ER应激激活通过ATF4依赖机构损害NIH3T3细胞中昼夜时钟和时钟控制基因的表达

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摘要

Endoplasmic reticulum (ER) stress and circadian clockwork signaling pathways mutually regulate various cellular functions, but the details regarding the cross-talk between these pathways in mammalian cells are unclear. In this study, whether perturbation of ER stress signaling affects the cellular circadian clockwork and transcription of clock-controlled genes was investigated in NIH3T3 mouse fibroblasts. An NIH3T3 cell model stably expressing luciferase (Luc) under the control of the Bmal1 clock gene promoter was established using a lentiviral system. Then, Luc activity was monitored in real-time to detect Bmal1-Luc oscillations. The ER stress activators thapsigargin (Tg) and tunicamycin (Tm) markedly reduced Bmal1-Luc oscillation amplitudes and induced phase delay shifts in NIH3T3 cells. Treatment with Tg/Tm activated ER stress signaling by upregulating GRP78, CHOP, ATF6, and ATF4 and simultaneously significantly decreased BMAL1 protein levels and inhibited the transcription of circadian clock (Bmal1, Per2, Nr1d1, and Dbp) and clock-controlled (Scad1, Fgf7, and Arnt) genes. 4-Phenylbutyric acid, an ER stress inhibitor, alleviated the transcriptional repression of the circadian clock genes and partially restored Bmal1-Luc oscillation amplitudes in Tg- or Tm-treated NIH3T3 cells. More importantly, knock-down of ATF4, but not ATF6, in Tg-treated NIH3T3 cells partially rescued Bmal1-Luc oscillation amplitudes and mRNA expression of the four circadian clock genes. Taken together, our study demonstrates that ER stress activation inhibits the transcription of circadian clock and clock-controlled genes via an ATF4-dependent mechanism.
机译:内质网(ER)压力和昼夜节律发条信号通路相互调节各种蜂窝功能,但是关于这些哺乳动物细胞中这些途径之间的交叉谈话的细节尚不清楚。在该研究中,ER应激信号传递的扰动是否会影响细胞昼夜节律钟表,并在NIH3T3小鼠成纤维细胞中研究了时钟控制基因的转录。使用慢病毒系统建立了在BMA1时钟基因启动子的控制下稳定表达荧光素酶(LUC)的NIH3T3细胞模型。然后,实时监测LUC活动以检测BMA1-LUC振荡。 ER应激活化剂Thapsigargin(Tg)和unicamycin(Tm)显着降低了BMA1-LUC振荡振荡幅度和NIH3T3细胞中的诱导相延迟移位。通过上调GRP78,CHOP,ATF6和ATF4并同时显着降低BMA11蛋白水平并抑制昼夜节日(BMAL1,PER2,NR1D1和DBP)和时钟控制(SCAD1, FGF7和ARNT)基因。 4-苯基丁酸,ER应激抑制剂,减轻了昼夜节日基因的转录抑制,在TG或TM处理的NIH3T3细胞中部分地恢复了BMA1-LUC振荡振荡。更重要的是,在TG处理的NIH3T3细胞中,ATF4的敲低,但不是ATF6部分地振动了四种昼夜钟表基因的BMA1-LUC振荡幅度和mRNA表达。我们的研究表明,ER应激激活通过ATF4依赖性机制抑制昼夜节点和时钟控制基因的转录。

著录项

  • 来源
    《Cellular Signalling》 |2019年第2019期|共13页
  • 作者单位

    Northwest A&

    F Univ Coll Vet Med Dept Clin Vet Med Yangling 752500 Shaanxi Peoples R China;

    Northwest A&

    F Univ Coll Vet Med Dept Clin Vet Med Yangling 752500 Shaanxi Peoples R China;

    Northwest A&

    F Univ Coll Vet Med Dept Clin Vet Med Yangling 752500 Shaanxi Peoples R China;

    Northwest A&

    F Univ Coll Vet Med Dept Clin Vet Med Yangling 752500 Shaanxi Peoples R China;

    Northwest A&

    F Univ Coll Vet Med Dept Clin Vet Med Yangling 752500 Shaanxi Peoples R China;

    Northwest A&

    F Univ Coll Vet Med Dept Clin Vet Med Yangling 752500 Shaanxi Peoples R China;

    Northwest A&

    F Univ Coll Vet Med Dept Clin Vet Med Yangling 752500 Shaanxi Peoples R China;

    Northwest A&

    F Univ Coll Vet Med Dept Clin Vet Med Yangling 752500 Shaanxi Peoples R China;

    Northwest A&

    F Univ Key Lab Anim Biotechnol Minist Agr Yangling 712100 Shaanxi Peoples R China;

    Northwest A&

    F Univ Coll Vet Med Dept Clin Vet Med Yangling 752500 Shaanxi Peoples R China;

  • 收录信息
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 细胞形态学;
  • 关键词

    Unfolded protein response; Circadian clock; Thapsigargin; Tunicamycin; 4-Phenylbutyric acid;

    机译:展开的蛋白质反应;昼夜节日;Thapsigargin;Tunicamycin;4-苯基丁酸;

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