DNA AND HYBRID STRUCTURE BY SOLUTION NMR

通过溶液 NMR 分析 DNA 和杂化结构

• 批准号: 6240437
• 负责人: BRIAN R REID
• 金额: $ 11.79万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-01-01 至 1997-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6240437
• 关键词: DNA; electronic spectra; endonuclease; gel mobility shift assay; molecular dynamics; molecular site; nuclear magnetic resonance spectroscopy; nucleic acid repetitive sequence; nucleic acid sequence; nucleic acid structure; solutions

A systematic study of nearest neighbor context effects on local base pair structure will be continued by completing the NMR-based solution structures of the 2 remaining structures in the CGNATNCG series and the three remaining structures in the CGNTANCG series. These 5 proposed sequences are the restriction sites for Bam H1, Sph I, Nhe I, Eco RV and Kpn I restriction endonucleases. The question of the critical size required to nucleate B-DNA in a B'-DNA environment will be elucidated by studying a series of duplexes of the type A5NnT5 (where n is varied from 1-4) in order to interpret gel retardation bending assays in oligomerized N5-iAiTiN5-i sequences. The structure of the bend at B-B' junctions and B'-B junctions will be studied in non-symmetrical dodecamers and in symmetrical hexadecamers in order to better define the helical axis of the flanking B-DNA segments. The structure and extent of bending will also be studied as a function of the flanking context sequence and the structural effects of G residues inserted into the B'oligo-A tract will be investigated. The structure of covalent junctions between DNA duplexes and hybrid DNA:RNA duplexes will be studied by NMR, especially retroviral junction sequences, and the ds DNA sequence at the right long-terminal-repeat of HIV-I will be investigated in an attempt to elucidate the structural basis for the ability of this LTR to integrate into the human genome. In addition to these goals, collaborative work on particular sequences of interest to other program laboratories will be carried out, and isotopic solid-state (DRAMA) atomic distance measurements will be undertaken with the Drobny lab to document structural peculiarities observed in our high-resolution structural studies.

对最近的邻居上下文对本地碱基对的影响的系统研究 通过完成基于NMR的解决方案将继续结构 CGNATNCG系列中其余两个结构的结构和 CGNTANCG系列中的剩余三个结构。 这5个提出 序列是BAM H1，SPH I，NHE I，ECO RV和 KPN I限制性核酸内切酶。 在B'-DNA中成核B-DNA所需的临界大小的问题 通过研究一系列的双工来阐明环境 键入A5NNT5（其中N与1-4不同）以解释凝胶 在寡聚的N5-iaitin5-I序列中进行延迟弯曲测定。 这 b-b'交界处和b'-b连接处的弯曲结构将是 在非对称的企业和对称的十六进制剂中进行了研究 为了更好地定义侧翼B-DNA段的螺旋轴。 弯曲的结构和程度也将作为功能研究 侧视上下文序列和g残基的结构效应 将研究插入B'Oligo-A区域。 DNA双链体与混合动力车之间的共价连接结构 DNA：RNA双链体将由NMR研究，尤其是逆转录病毒连接 序列，以及右长末端重复的DS DNA序列 HIV-I将进行研究，以阐明结构 该LTR能够整合到人类基因组中的能力的基础。 除了这些目标，还要在特定序列上进行协作工作 其他计划实验室感兴趣的人将进行，并 同位素固态（戏剧）原子距离测量将为 与Drobny Lab一起进行记录的结构特征 在我们的高分辨率结构研究中观察到。