• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Molecular and Cellular Biology >Identification of Cyclin D3 as a Direct Target of E2A Using DamID
【24h】

Identification of Cyclin D3 as a Direct Target of E2A Using DamID

机译：使用DamID识别Cyclin D3作为E2A的直接靶标

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

摘要

The transcription factor E2A can promote precursor B cell expansion, promote G1 cell cycle progression, and induce the expressions of multiple G1-phase cyclins. To better understand the mechanism by which E2A induces these cyclins, we characterized the relationship between E2A and the cyclin D3 gene promoter. E2A transactivated the 1-kb promoter of cyclin D3, which contains two E boxes. However, deletion of the E boxes did not disrupt the transactivation by E2A, raising the possibility of indirect activation via another transcription factor or binding of E2A to non-E-box DNA elements. To distinguish between these two possibilities, promoter occupancy was examined using the DamID approach. A fusion construct composed of E2A and the Escherichia coli DNA adenosine methyltransferase (E47Dam) was subcloned in lentivirus vectors and used to transduce precursor B-cell and myeloid progenitor cell lines. In both cell types, specific adenosine methylation was identified at the cyclin D3 promoter. Chromatin immunoprecipitation analysis confirmed the DamID findings and localized the binding to within 1 kb of the two E boxes. The methylation by E47Dam was not disrupted by mutations in the E2A portion that block DNA binding. We conclude that E2A can be recruited to the cyclin D3 promoter independently of E boxes or E2A DNA binding activity.
机译：转录因子E2A可以促进前体B细胞的扩增，促进G 1 细胞周期的进程，并诱导多个G 1 期细胞周期蛋白的表达。为了更好地理解E2A诱导这些细胞周期蛋白的机制，我们表征了E2A和细胞周期蛋白D3基因启动子之间的关系。 E2A激活了细胞周期蛋白D3的1-kb启动子，该启动子包含两个E盒。然而，E盒的缺失并未破坏E2A的反式激活，增加了通过另一种转录因子间接激活或E2A与非E盒DNA元件结合的可能性。为了区分这两种可能性，使用DamID方法检查了启动子的占用。将由E2A和大肠杆菌 DNA腺苷甲基转移酶（E47Dam）组成的融合构建体亚克隆到慢病毒载体中，并用于转导前体B细胞和骨髓祖细胞系。在这两种细胞类型中，在细胞周期蛋白D3启动子处均鉴定出特定的腺苷甲基化。染色质免疫沉淀分析证实了DamID的发现并将结合定位于两个E盒的1kb内。 E47Dam的甲基化不会被阻断DNA结合的E2A部分中的突变所破坏。我们得出的结论是，E2A可以独立于E盒或E2A DNA结合活性而募集到细胞周期蛋白D3启动子。

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