• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Molecular and Cellular Biology >An inducible 50-kilodalton NF kappa B-like protein and a constitutive protein both bind the acute-phase response element of the angiotensinogen gene.
【24h】

An inducible 50-kilodalton NF kappa B-like protein and a constitutive protein both bind the acute-phase response element of the angiotensinogen gene.

机译：可诱导的50-Kaldaltonton NF kappa B样蛋白和组成型蛋白都结合血管紧张素原基因的急性期反应元件。

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The rat angiotensinogen gene is induced in the course of the hepatic acute-phase response. We demonstrate that monocyte conditioned medium can stimulate transcription of a stably introduced reporter construct driven by 615 base pairs of the angiotensinogen 5'-flanking sequence, as well as the endogenous gene, in Reuber H35 cells. Point mutations of a cis-acting element, located 545 base pairs from the transcription start site and sharing sequence identity with known nuclear factor kappa B (NF kappa B)-binding sites, led to loss of cytokine inducibility. When cloned upstream of a minimal promoter, this cis-acting element imparted transcriptional inducibility by monocyte conditioned medium, interleukin-1, and tumor necrosis factor on a luciferase reporter gene in HepG2 cells. Two distinct proteins bound this element in vitro: a heat-stable, constitutively present, hepatic nuclear protein that gave rise to a DNase I-protected footprint covering the functionally defined element; and a binding protein of different mobility, induced by monocyte conditioned medium, which also recognized the NF kappa B-binding site of the murine kappa light-chain enhancer. UV cross-linking showed this inducible protein to have an apparent molecular mass of 50 kilodaltons, similar to that described for NF kappa B and distinct from the constitutively present protein that was shown by Southwestern (DNA-protein) blot to have a molecular mass of 32 kilodaltons. Methylation interference analysis showed that the induced species made contact points with guanine residues in the NF kappa B consensus sequence typical of NF kappa B. Induction of this binding activity did not require new protein synthesis, and 12-O-tetradecanoylphorbol-13-acetate could mimic the induction by cytokines. We thus provide direct evidence for involvement of NF kappa B or a similar factor in the hepatic acute-phase response and discuss the potential role of the presence of a constitutive nuclear factor binding the same cis element.
机译：大鼠血管紧张素原基因在肝急性期反应过程中被诱导。我们证明单核细胞条件培养基可以刺激Reuber H35细胞中由血管紧张素原5'侧翼序列的615个碱基对以及内源基因驱动的稳定引入的报告基因构建体的转录。顺式作用元件的点突变位于距转录起始位点545个碱基对的位置，并且与已知的核因子κB（NF kappa B）结合位点具有相同的序列同一性，导致细胞因子诱导性的丧失。当克隆到最小启动子的上游时，该顺式作用元件通过单核细胞条件培养基，白介素-1和肿瘤坏死因子在HepG2细胞中的萤光素酶报告基因上赋予转录诱导性。两种不同的蛋白质在体外结合了该元素：一种热稳定的，组成性存在的肝核蛋白，其产生了由DNase I保护的覆盖功能上定义的元素的印迹;以及由单核细胞条件培养基诱导的具有不同迁移率的结合蛋白，该结合蛋白也能识别鼠κ轻链增强子的NFκB结合位点。 UV交联表明，这种诱导型蛋白质的表观分子量为50道尔顿，与NFκB的分子量相似，并且不同于西南蛋白质（DNA-蛋白质）印迹显示的组成型蛋白质的分子量为32公里。甲基化干扰分析表明，诱导的物种与NFκB共有的NFκB共有序列中的鸟嘌呤残基接触。诱导这种结合活性不需要新的蛋白质合成，而12-O-十四烷酰佛波醇13-乙酸盐可以模拟细胞因子的诱导。因此，我们提供了NFκB或类似因子参与肝急性期反应的直接证据，并讨论了结合相同顺式元件的组成性核因子的潜在作用。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号