Core 1 - NMR

核心 1 - 核磁共振

• 批准号: 10245108
• 负责人: MICHAEL FINLEY SUMMERS
• 金额: $ 47.61万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-17 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10245108
• 关键词: Antiviral Agents; Base Pairing; Chemicals; Collection; Computer software; Cryoelectron Microscopy; Databases; Deposition; Detection; Development; Funding; HIV; Hybrids; Institution; Isotope Labeling; Isotopes; Label; Ligands; Measurement; Measures; Methodology; Methods; Molecular Conformation; Mutagenesis; Nucleotides; Parents; Physiologic pulse; Preparation; Proteins; RNA; RNA Editing; RNA analysis; Reagent; Research Personnel; Ribonucleosides; Running; Services; Signal Transduction; Site; Structure; T7 RNA polymerase; United States National Institutes of Health; Validation; Work; base; computerized data processing; data acquisition; improved; innovation; member; millisecond; novel; novel strategies; predictive tools; prevent; screening services; small molecule; structural biology; tripolyphosphate

Core 1: NMR Summary The NMR Core includes investigators at 6 institutions who are developing isotopically labeled ribonucleoside triphosphate (rNTPs) reagents and RNA labeling methodologies (Dayie, Williamson), and NMR data acquisition/assignment methodologies (Al-Hashimi, Summers, Marchant, Tolbert, Dayie, Johnson, Bax), to facilitate NMR studies of large RNAs. The NMR Core also interacts extensively with the Case and Johnson labs of the Computational Core, and with the Chiu, Zheng, and Pornillos labs of the cryoEM Core, to facilitate development of hybrid methodologies and force fields that enable structural and dynamical studies of larger HIV RNAs. Hybrid NMR/SAXS methods are also being developed and evaluated (Tolbert, Case, Dayie). The NMR Core also provides screening services to assist CRNA members in the identification of viable new RNA targets for structural analysis, and to support efforts by Hargrove to identify RNA-targeting antivirals. Over the past four years, the NMR Core has contributed to several technical innovations and discoveries including: (1) development of a novel fragmentation/annealing-based approach (fr-RNA) for detection and signal assignment of long-range secondary structures; (2) development of improved nucleotide-specific partial deuteration strategies (2H-editing) for NMR signal assignment; (3) development of improved methods detecting and preventing self-templated run-on during T7 RNA polymerase dependent synthesis; (4) development of a novel NMR method for unambiguous identification of intermolecular base pairing in large RNAs; (5) development of a novel NMR/mutagenesis method for measuring the rate of formation of intermolecular base pairs; (6) development of new tools for predicting and validating 1H NMR signal assignments by analysis of depositions in the BioMagResBank database; (7) contributed to the development of a new software package for rapid NMR data processing, and semi-automated RNA signal assignment; (8) development of new pulse sequences for rapid collection of isotopically edited RNA NMR spectra; and for the measurement and characterization of micro-to-millisecond conformational exchange in RNA; (9) development of new 2H, 13C, and 15N-labeled reagents to facilitate structural and dynamical studies of large RNAs; and (10) conducted service work for members of the CRNA (Beemon, Bieniasz, Hargrove, Boris-Lawrie, Parent, Heng), as well as for other NIH funded centers (James Hurley, HARC, Berkeley), several of which led to new structural projects. The NMR Core will continue to push the size limits of NMR and will support studies of new protein and RNA targets identified by the CRNA and other NIH funded groups. The methodological innovations that have been developed are generally applicable to the rapidly expanding field of RNA structural biology.

核心1：核磁共振 摘要 核磁共振核心包括6个机构的研究人员，他们正在开发同位素标记的核糖核苷 三磷酸(RNTPs)试剂和RNA标记方法(Daie，Williamson)和核磁共振数据 收购/转让方法(Al-Hashimi、Summers、Marchant、Tolbert、Dayie、Johnson、Bax)，至 促进大RNA的核磁共振研究。核磁共振核心还与凯斯和约翰逊进行广泛的互动 计算核心实验室，以及CryoEM核心的Chiu、Zeng和Pornillos实验室，以促进 开发混合方法和力场，使更大规模的结构和动力学研究成为可能 HIV核糖核酸。还在开发和评估核磁共振/小角X射线散射混合方法(Tolbert、Case、Daie)。这个 核磁共振核心还提供筛选服务，以协助CRNA成员识别可行的新RNA 结构分析的目标，并支持哈格罗夫为确定以RNA为靶标的抗病毒药物所作的努力。超过了 在过去的四年里，核磁共振核心为几项技术创新和发现做出了贡献，包括：(1) 一种新的基于碎片/退火法的检测和信号分配方法(fr-RNA)的发展 长程二级结构；(2)改进的核苷酸特异性部分重氢反应的发展 核磁共振信号赋值的策略(2H编辑)；(3)改进的检测和检测方法的发展 防止T7RNA聚合酶依赖合成过程中的自模板跑动；(4)一种新型的 核磁共振方法用于明确鉴定大RNA中分子间碱基配对；(5) 测定分子间碱基对形成速率的核磁共振/诱变新方法(6) 通过沉积分析预测和验证~1H核磁共振信号赋值的新工具的开发 在BioMagResBank数据库中；(7)协助开发了一个用于快速核磁共振的新软件包 数据处理和半自动RNA信号分配；(8)开发新的脉冲序列 快速收集以同位素编辑的RNA核磁共振谱；以及用于测量和表征 RNA中微秒到毫秒的构象交换；(9)新的2H、13C和15N标记的发展 促进大型核糖核酸的结构和动力学研究的试剂；以及(10)为 CRNA成员(Beemon、Bieniasz、Hargrove、Boris-Lawrie、Parent、Heng)以及其他NIH成员 资助的中心(詹姆斯·赫尔利、HARC、伯克利)，其中几个导致了新的结构项目。核磁共振 CORE将继续推动核磁共振的大小限制，并将支持新的蛋白质和RNA靶标的研究 由CRNA和其他NIH资助的组织确定。一直以来的方法论创新 所开发的方法普遍适用于迅速发展的RNA结构生物学领域。