CHARACTERIZATION OF THE GENE PRODUCTS OF THE ETS LOCUS

ETS 基因座基因产物的表征

• 批准号: 3838373
• 负责人: T S PAPAS
• 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/3838373
• 关键词: DNA binding protein; gel mobility shift assay; gene expression; human tissue; nucleic acid probes; nucleic acid sequence; oligonucleotides; oncogenes; oncoproteins; protein purification; protein sequence; protooncogene

The ETS1 protein purified from CEM cells was used to select its optimum DNA binding sequence (pu) G/C C/a G G A A G T/c (py). The sequence CCGGAAGT (ETS1-3) was preferred 5:1 over CAGGAAGT (PEA3). Quantitative electrophoretic mobility shift assays (EMSA) indicate that the purified ETS1 protein binds to either ETS1-3 or PEA3 oligonucleotide probes with high affinity, and that the purified ETS1 has different binding capacities for ETS1-3 and PEA3 oligonucleotide probes. The ETS1 protein binds 2-5 times more to the ETS1-3 than the PEA3 probe. Competitive binding experiments show that the ETS1-3 and PEA3 probes effectively compete for the binding of ETS1-3. However, change of the core DNA-binding sequence from GGAA to AGAA eliminates competition. The human ETS1 protein selected the same DNA sequence from a mixture of random oligonucleotides, as did the Drosophila E74A protein (one of the most divergent members of the ETS family), strongly suggesting that all proteins containing the ETS 85 amino acid domain (sequences which define the ETS family) will bind to the same sequence.

从CEM细胞中纯化的ETS1蛋白进行筛选