Chen J H, Wright C D
Department of Experimental Pediatrics, University of Texas M.D. Anderson Cancer Center, Houston 77030.
Oncogene. 1993 Dec;8(12):3375-83.
The promoter of the human ETS1 gene contains binding site motifs for transcription factors Sp1, Ap1, Ap2, ETS, PEA3 and Oct. It has been shown previously that Ap1, Ap2, and ETS are positive regulators for the transcription of the ETS1 gene and that the ETS1 gene is autoregulated. In addition, two regions on the ETS1 promoter that contain negative regulatory sequences have also been identified. In this study, GST/PEA3 and GST/ets1 fusion proteins were used in gel mobility assays to analyse the interaction between these transcription factors with the binding motifs of PEA3 and ETS in the ETS1 promoter. Promoter constructs in which the binding site motifs are mutated were used to examine the effect of mutation on the activities of transcription factors PEA3, ets1 and Oct. Reporter plasmids containing different deletion mutants of the ETS1 promoter were used to examine the effect of different transcription factors (PEA3, Oct 1 and Oct 2) upon the expression of the ETS1 gene. The results of these studies indicate that PEA3 is a strong positive regulator of the ETS1 promoter and that other transcription factors increase the activity of the ETS1 promoter in an additive rather than synergistic fashion.
人类ETS1基因的启动子包含转录因子Sp1、Ap1、Ap2、ETS、PEA3和Oct的结合位点基序。先前已经表明，Ap1、Ap2和ETS是ETS1基因转录的正调节因子，并且ETS1基因是自我调节的。此外，还鉴定出了ETS1启动子上两个含有负调节序列的区域。在本研究中，利用GST/PEA3和GST/ets1融合蛋白进行凝胶迁移试验，以分析这些转录因子与ETS1启动子中PEA3和ETS的结合基序之间的相互作用。使用结合位点基序发生突变的启动子构建体来检测突变对转录因子PEA3、ets1和Oct活性的影响。使用含有ETS1启动子不同缺失突变体的报告质粒来检测不同转录因子（PEA3、Oct 1和Oct 2）对ETS1基因表达的影响。这些研究结果表明，PEA3是ETS1启动子的强正调节因子，并且其他转录因子以累加而非协同的方式增加ETS1启动子的活性。
