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Identification of regulatory elements within the minimal promoter region of the human endogenous ERV9 proviruses: accurate transcription initiation is controlled by an Inr-like element.

机译:鉴定人类内源ERV9原病毒最小启动子区域内的调控元件:准确的转录起始受Inr样元件控制。

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摘要

ERV9 is a low repeated family of human endogenous retroviral elements whose expression is mainly detectable in undifferentiated embryonal carcinoma NT2/D1 cells. In this report we have analyzed the minimal promoter region located within the ERV9 LTR. Using the transient CAT expression assay we have identified the minimal promoter region, which includes sequences spanning from -70 to +6 relative to the major transcription start site. Deletion analysis, primer extension mapping of the transcription start sites and DNA-protein interactions assays have allowed us to define two important regions within the ERV9 minimal promoter. One region located between -70 to -39 acts as a transcriptional activating sequence and contains an Sp 1 binding site. The second region from -7 to +6, which resembles an initiator element (Inr), was necessary for the correct transcription start site utilization, and binds to a regulatory protein. Cross-competition experiments using various Inr elements have indicated that the protein that binds to the ERV9 Inr element can be competed by the HIV-1 and TdT Inr sequences.
机译:ERV9是人类内源性逆转录病毒元件的低重复家族,其表达主要在未分化的胚胎癌NT2 / D1细胞中检测到。在此报告中,我们分析了位于ERV9 LTR内的最小启动子区域。使用瞬时CAT表达测定,我们已经确定了最小启动子区域,其中包括相对于主要转录起始位点从-70到+6的序列。缺失分析,转录起始位点的引物延伸图谱和DNA-蛋白质相互作用测定使我们能够在ERV9最小启动子内定义两个重要区域。位于-70至-39之间的一个区域充当转录激活序列,并包含Sp 1结合位点。类似于启动子元件(Inr)的从-7到+6的第二个区域对于正确转录起始位点的利用是必需的,并与调节蛋白结合。使用各种Inr元素的交叉竞争实验表明,与ERV9 Inr元素结合的蛋白质可以与HIV-1和TdT Inr序列竞争。

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