TETRALOOP RECEPTOR RNA STRUCTURAL INVESTIGATION

四环受体 RNA 结构研究

• 批准号: 8361194
• 负责人: Samuel E Butcher
• 金额: $ 1.36万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361194
• 关键词: Bacteriophage Pf1; Binding; COSY; Functional RNA; Funding; Grant; Investigation; Kinetics; Measures; Mediating; NOESY; National Center for Research Resources; Principal Investigator; RNA; Research; Research Infrastructure; Residual state; Resources; Sampling; Source; Structure; TOCSY; Temperature; United States National Institutes of Health; Work; cost; receptor; research study

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We are investigating the structure of the free tetraloop receptor. The tetraloop receptor is a commonly occurring motif that mediates tertiary folding of large, non-coding RNAs. We are investigating the tetraloop receptor structure as a function of temperature and ionic conditions. Previous work shows that the tetraloop receptor undergoes a conformational change upon tetraloop binding. We are also investigating the kinetics of this conformational change by NMR. NMR experiments that we will need to collect include: 1D (1H), 2D NOESY, 2D 1H-15N HSQC (normal and TROSY enhanced), 2D 1H-15N HNN COSY, 2D 1H-13C HSQC (normal and TROSY enhanced), 2D NOESY, 2D 1H,13C filter/edit NOESY, 3D 1H(13C)1H NOESY-HSQC, 3D 1H(13C)1H HCCH TOCSY and 3D 1H(13C)1H HCCH COSY. Residual dipolar couplings will be measured for samples in both isotropic and partially aligned states (via addition of Pf1 phage).

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 我们正在研究游离四环受体的结构。 四环受体是一种常见的基序，介导大非编码 RNA 的三级折叠。 我们正在研究四环受体结构作为温度和离子条件的函数。 先前的工作表明，四环受体在四环结合时会发生构象变化。 我们还通过核磁共振研究了这种构象变化的动力学。 我们需要收集的 NMR 实验包括：1D (1H)、2D NOESY、2D 1H-15N HSQC（正常和 TROSY 增强）、2D 1H-15N HNN COSY、2D 1H-13C HSQC（正常和 TROSY 增强）、2D NOESY，2D 1H，13C 过滤/编辑 NOESY，3D 1H(13C)1H NOESY-HSQC、3D 1H(13C)1H HCCH TOCSY 和 3D 1H(13C)1H HCCH COSY。将测量各向同性和部分对齐状态（通过添加 Pf1 噬菌体）的样品的残余偶极耦合。