DNA MINOR GROOVE BINDING POLYAMIDES

DNA 小沟结合聚酰胺

• 批准号: 6626692
• 负责人: TERRY A BEERMAN
• 金额: $ 32.79万
• 依托单位: ROSWELL PARK CANCER INSTITUTE CORP
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-01-09 至 2004-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6626692
• 关键词: DNA; DNA footprinting; amides; chemical binding; genetic promoter element; neoplasm /cancer genetics; polymers; protooncogene; transcription factor; transcription termination

DESCRIPTION (verbatim from the applicant's abstract): This research program seeks to devise new strategies to rationally inhibit gene expression mediated by transcription factors (TFs). The strategy is to design modular polyamide DNA minor binding agents that potently and specifically inhibit TF binding to gene promoters and disrupt gene expression. For this purpose, a representative of the Ets oncogenic protein family Elk-1, which binds a serum response element (SRE) contained within the c-fos promoter, was chosen as a model TF. Interference with the binding of Elk-1 to the SRE will be quantitatively measured by mobility shift assays, to assess a polyamide's ability to inhibit TF/DNA complexes. Quantitative footprint analysis will be used to determine where polyamides bind on the SRE promoter element and also the strength of the binding. Polyamides that are potent and specific inhibitors of Elk-1/SRE complex formation will be evaluated for their ability to inhibit transcriptional activation from the c-fos promoter in cell-free and cellular assays. Promising agents will be evaluated for their ability to interfere with a neoplastic phenotype associated with c-fos gene expression. In parallel, polyamides will be optimized for cellular activity while maintaining target specificity. Specific aims: Aim1. Identification of polyamide DNA minor groove binding agents that interfere with the binding of Elk-1 to the c-fos promoter under cell-free conditions. Aim2. Effects of polyamide DNA minor groove binding agents on Elk-1 mediated gene expression of the c-fos promoter under cell-free conditions. Aim3. Effects of polyamide DNA minor groove binding agents on Elk-1 mediated gene expression of the c-fos promoter in intact cells.

描述（逐字研究申请人的摘要）：该研究计划 试图制定新策略以合理抑制介导的基因表达 通过转录因子（TFS）。该策略是设计模块化聚酰胺DNA 次要结合剂，有效，专门抑制TF与基因的结合 启动子和破坏基因表达。为此目的， ETS致癌蛋白家族ELK-1，结合血清反应元件 （SRE）包含在C-FOS启动子中，选择作为Model TF。 干扰ELK-1与SRE的结合将是定量的 通过移动性转移测定法测量，以评估聚酰胺的抑制能力 TF/DNA复合物。定量足迹分析将用于确定 聚酰胺结合在SRE启动子元件上以及强度 结合。 ELK-1/SRE的有效抑制剂的聚酰胺 将评估复合形成的抑制能力 无细胞和细胞中C-FOS启动子的转录激活 测定。有希望的代理人将对他们干扰的能力进行评估 与C-FOS基因表达相关的肿瘤表型。并联， 聚酰胺将用于细胞活性，同时保持靶标 特异性。具体目的：AIM1。鉴定聚酰胺DNA小凹槽 结合剂，干扰ELK-1与C-FOS启动子的结合 在无细胞条件下。 AIM2。聚酰胺DNA小凹槽结合的影响 C-FOS启动子在无细胞下的ELK-1介导的基因表达的药物 状况。 AIM3。聚酰胺DNA小凹槽结合剂对ELK-1的影响 完整细胞中C-FOS启动子的介导基因表达。