NONCOVALENT AND COVALENT DNA MODIFICATION

非共价和共价 DNA 修饰

基本信息

批准号：
3071536
负责人：
JOHN W SUGGS
金额：
$ 5.62万
依托单位：
BROWN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1984
资助国家：
美国
起止时间：
1984-07-01 至 1989-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3071536
关键词：
DNA chemical bond chemical structure function circular dichroism conformation covalent bond molecular site netropsin nuclear magnetic resonance spectroscopy nucleic acid sequence nucleic acid structure oligonucleotides oncogenes phosphorus polynucleotides synthetic nucleotide thiols

项目摘要

Our long range goal is to design and synthesize organic molecules which bind to specific DNA sequences and to covalently modify specific DNA sites. Our first immediate subgoal is to characterize the solution structure of a new DNA conformation we have discovered, a dinucleotide repeat structure which is induced by binding certain steroid diamines to poly(dA-dT). Using NMR and CD to probe DNA conformation we will synthesize new ligands to see what structural features of the steroid diamine promote conformational change in poly(dA-dT). We will chemically synthesize small DNA oligomers to discover what sequences can exist in this new conformation, using NMR as a conformational probe of the DNA-drug complex. We will make DNA with phosphorothioate groups in the backbone to study backbone conformations in detail. Such thio-DNAs will also be used to covalently attach groups to the DNA. We also propose to synthesize new linked drugs containing netropsin and anthramycin fragments. These linked groove binders should exhibit unprecedented sequence selectivity by a synthetic molecule. We will assay selectivity by the drug's differential inhibition of site selectivity of particular restriction enzymes. This work should eventually lead to molecules which can act as repressors of specific genes (incluing oncogenes). It will also provide a way to introduce specific covalent lesions into specific DNA sequences, providing a clean experimental system for determining the mutational properties of DNA backbone alkylation.

我们的长期目标是设计和合成有机分子结合特定 DNA 序列并共价修饰特定 DNA 网站。我们的第一个直接子目标是描述解决方案的特征我们发现的一种新的DNA构象的结构，一种二核苷酸通过与某些类固醇二胺结合而诱导的重复结构聚（dA-dT）。使用 NMR 和 CD 来探测 DNA 构象，我们将合成新配体，看看类固醇二胺促进了哪些结构特征聚（dA-dT）的构象变化。我们将化学合成小的 DNA寡聚物来发现这个新的序列中可以存在什么序列构象，使用 NMR 作为 DNA-药物复合物的构象探针。我们将制作主链带有硫代磷酸酯基团的DNA来进行研究详细的骨干构象。此类 thio-DNA 还将用于将基团共价连接到 DNA 上。我们还建议合成新的含有 netropsin 和 anthramycin 片段的连接药物。这些链接的凹槽结合物应表现出前所未有的序列选择性合成分子。我们将通过药物的差异来测定选择性抑制特定限制性酶的位点选择性。这项工作最终应该会产生可以充当阻遏物的分子特定基因（包括癌基因）。它还将提供一种方法将特定的共价损伤引入特定的 DNA 序列中，提供一个干净的实验系统，用于确定突变特性 DNA骨架烷基化。