DNA MINOR GROOVE BINDING POLYAMIDES

描述（来自申请人摘要的逐字记录）：本研究项目 试图设计新的策略来合理地抑制基因表达介导的 转录因子（TF）。该策略是设计模块化聚酰胺DNA 有效和特异性抑制TF与基因结合的次要结合剂 启动子并破坏基因表达。为此， Ets致癌蛋白家族Elk-1，其结合血清应答元件 (SRE)包含在c-fos启动子内的基因，被选为模型TF。 将定量测定Elk-1与SRE结合的干扰 通过迁移率变动测定测量，以评估聚酰胺抑制 TF/DNA复合物。定量足迹分析将用于确定 其中聚酰胺结合在SRE促进剂元件上， 约束力作为Elk-1/SRE的有效和特异性抑制剂的聚酰胺 将评价复合物形成抑制 c-fos启动子在无细胞和细胞中的转录激活 测定。有前途的代理人将评估他们的能力，干扰 与c-fos基因表达相关的肿瘤表型。同时， 聚酰胺将针对细胞活性进行优化，同时保持目标 的特异性具体目标：目标1。聚酰胺DNA小沟的鉴定 干扰Elk-1与c-fos启动子结合的结合剂 在无细胞条件下。目标2。聚酰胺DNA小沟结合的影响 药物对Elk-1介导的c-fos启动子基因表达的影响 条件目标3。聚酰胺类DNA小沟结合剂对Elk-1的作用 介导的c-fos启动子在完整细胞中的基因表达。

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.