• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Molecular and Cellular Biology >Cell-specific transcriptional regulation and reactivation of galectin-1 gene expression are controlled by DNA methylation of the promoter region.
【24h】

Cell-specific transcriptional regulation and reactivation of galectin-1 gene expression are controlled by DNA methylation of the promoter region.

机译：galectin-1基因表达的细胞特异性转录调控和再激活受启动子区域DNA甲基化的控制。

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

摘要

The galectin-1 gene is developmentally regulated gene whose activity is strongly modulated during cell differentiation and transformation. We have previously shown that galectin-1 promoter constructs are highly active when transiently transfected in cells both expressing and not expressing the endogenous gene and that the basal activity is determined by a small region encompassing the transcription start site (from positions -50 to +50). We have now investigated the role of DNA methylation in galectin-1 gene expression. Southern blot analysis with HpaII and MspI endonucleases and sodium bisulfite analysis of genomic DNA from expressing and nonexpressing cell lines and cell hybrids showed a close correlation between gene activity and demethylation of the 5' region of the galectin-1 gene. We found that the galectin-1 promoter region is fully methylated, at every CpG site on both strands, in nonexpressing differentiated rat liver (FAO) and thyroid (PC C13) cells and unmethylated in the expressing undifferentiated liver (BRL3A) and thyroid transformed (PC myc/raf) cell lines. In addition, reactivation of the silent FAO alleles in FAO-human osteosarcoma (143tk-) hybrid cells is accompanied by a complete demethylation of the promoter region. Finally, when galectin-1 chloramphenicol acetyltransferase (CAT) promoter constructs were methylated in vitro by SssI methylase at every cytosine residue of the CpG doublets and transfected into mouse fibroblasts, the transcription of the CAT reporter gene was strongly inhibited.
机译：galectin-1基因是发育受调控的基因，其活性在细胞分化和转化过程中受到强烈调节。先前我们已经表明，当在表达和不表达内源基因的细胞中瞬时转染时，galectin-1启动子构建体均具有很高的活性，并且基础活性由包含转录起始位点的小区域（从-50到+50位置）决定）。我们现在研究了半乳糖凝集素-1基因表达中DNA甲基化的作用。用HpaII和MspI核酸内切酶进行的Southern印迹分析以及表达和不表达的细胞系和细胞杂种的基因组DNA的亚硫酸氢钠分析表明，基因活性与galectin-1基因5'区域的去甲基化密切相关。我们发现，在未表达的分化大鼠肝脏（FAO）和甲状腺（PC C13）细胞中，两条链上每个CpG位点的galectin-1启动子区均被完全甲基化，而在未表达的未分化肝脏（BRL3A）和已转化的甲状腺中，半乳糖凝集素1启动子区域未甲基化（ PC myc / raf）细胞系。此外，在粮农组织-人骨肉瘤（143tk-）杂交细胞中沉默的粮农组织等位基因的重新激活伴随着启动子区域的完全脱甲基。最后，当半乳糖凝集素-1氯霉素乙酰转移酶（CAT）启动子构建体在体外通过SssI甲基化酶在CpG双峰的每个胞嘧啶残基处甲基化并转染到小鼠成纤维细胞中时，CAT报告基因的转录受到强烈抑制。

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