TRANSCRIPTION REGULATION OF THE HUMAN ETS-2 ONCOGENE

人类 ETS-2 癌基因的转录调控

• 批准号: 3874741
• 负责人: D K WATSON
• 金额: --
• 依托单位: DIVISION OF CANCER EPIDEMIOLOGY AND GENETICS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3874741
• 关键词: DNA binding protein; chemical binding; fusion gene; gel electrophoresis; gene frequency; genetic mapping; genetic promoter element; genetic regulatory element; genetic transcription; human population genetics; molecular cloning; neoplastic cell culture for noncancer research; nucleic acid sequence; oncogenes; point mutation; purine nucleosides; purines; pyrimidine nucleosides; pyrimidines; site directed mutagenesis; transcription factor; transfection

We have previously identified the region which is required for the ETS2 transcription. We used the minimal ETS2 promoter region to identify factors that are involved in ETS2 transcriptional regulation. Gel mobility shift and methylation interference assays reveal that at least five distinct protein complexes interact with the region surrounding the major transcription initiation site. None of the interactions involve known proteins, as indicated by the inability to complete the effect with target sequences by known transcription factors (Sp1, AP1, AP2, AP3, NF1). Two of the DNA protein complexes involve the GC element with dyad symmetry which we previously identified to exist in other genes. Three other complexes have been localized to the region immediately 3' of the major transcription initiation points; one of these complexes is lymphoid specific and the others are ubiquitous (detected in every nuclear extract tested). Point mutagenesis of the DNA binding sites and subsequent determination of the promoter activity reveals that the integrity of the sites is required for proper ETS2 MRNA transcription. The identification of the DNA binding sites on the ETS2 promoter will allow the isolation and characterization of the genes involved in the ETS2 transcription unit.

我们之前已经确定了ETS 2所需的区域 转录。我们使用最小的ETS 2启动子区域来鉴定 参与ETS 2转录调控的基因。凝胶迁移率变动 和甲基化干扰试验表明，至少有五个不同的 蛋白质复合物与主要的 转录起始位点。没有一个互动涉及已知的 蛋白质，如通过不能完成与靶的作用所指示的 已知的转录因子（Sp1，AP 1，AP 2，AP 3，NF 1）。两 DNA蛋白质复合物涉及具有二分体对称性的GC元件， 我们之前发现存在于其他基因中。其他三个复合体 已经定位于紧邻主要转录的3'的区域 起始点;这些复合物之一是淋巴特异性的， 其他的则是普遍存在的（在每一种测试的核提取物中检测到）。点 DNA结合位点的诱变和随后的 启动子活性揭示了位点的完整性是启动子活性所必需的。 ETS 2 mRNA转录。DNA结合位点的鉴定 ETS 2启动子上的突变将允许分离和表征 参与ETS 2转录单位的基因。