HELICAL RNA, RNA:DNA HYBRIDS AND DNA-RNA JUNCTIONS

螺旋 RNA、RNA:DNA 杂交体和 DNA-RNA 连接

• 批准号: 3301823
• 负责人: BRIAN R REID
• 金额: $ 18.85万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-07-01 至 1994-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3301823
• 关键词: DNA; RNA; chemical cleavage; fusion gene; gene duplication; gene expression; gene redundancy; genetic promoter element; messenger RNA; molecular shape; nuclear magnetic resonance spectroscopy; nucleic acid sequence; transfer RNA

RNA duplexes, RNA:DNA hybrid duplexes and DNA-RNA duplex junctions are involved in many important steps in gene expression including replication, transcription and translational control. Relatively little is known about the structure of these biologically important duplexes, largely due to the difficulty of synthesizing the large amounts of defined sequence RNA molecules required for detailed structural studies (NMR, x-ray crystallography). We have circumvented this bottleneck and have recently synthesized 20-50 mg quantities of RNA chains with defined sequences. We now propose to carry out extensive high-resolution NMR studies on the solution structure of r:r duplexes and to compare their structure to d:d duplexes of the same Studies will also be carried out on d:r hybrid duplexes and d-r-d:d-r-d chimeric duplexes containing A-B junctions. Initial insights into structure will be obtained from complete proton assignments, chemical shift changes, and interpretation of changes in coupling constants. More detailed later structural studies will involve time-dependent NOESY studies, interproton distance measurements and ab initio generation of three-dimensional structures consistent with these distances using distance geometry algorithms. The results should shed a great deal of light on the peculiarities of RNA structure, the control of transcription of DNA information into RNA information (and vice-versa) and the apparent need for RNA primers at replication forks and in the initiation of DNA replication.

RNA双链体，RNA：DNA混合双链体和DNA-RNA双链连接 参与了基因表达的许多重要步骤，包括 复制，转录和翻译控制。 相对地 关于这些生物学重要的结构知之甚少 双工，很大程度上是由于难以合成大的 详细所需的定义序列RNA分子量 结构研究（NMR，X射线晶体学）。 我们有 绕过这种瓶颈，最近合成了20-50 具有定义序列的RNA链的Mg量。 我们现在建议进行广泛的高分辨率NMR研究 关于R：R双链体的溶液结构，并比较其 同一研究的D：D的双链体的结构也将被携带 在D：R混合双链体和D-R-D：D-R-D Chimeric双工 包含A-B连接。 对结构的初步见解将从完整中获得 质子分配，化学转移变化和解释 耦合常数的变化。 更详细的以后结构 研究将涉及时间依赖于时间的NOESY研究，Intersoton 距离测量和三维的初始生成 使用距离几何形状与这些距离一致的结构 算法。 结果应该对特殊性阐明 RNA结构，DNA信息转录的控制 进入RNA信息（反之亦然），明显的RNA需求 复制叉和DNA的启动时的底漆 复制。