TETRALOOP RECEPTOR RNA STRUCTURAL INVESTIGATION

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 我们正在研究游离的四环素受体的结构。Tetraloop受体是一种常见的基序，介导大的非编码RNA的三级折叠。我们正在研究四环素受体结构随温度和离子条件的变化。以前的工作表明，四环受体在四环结合时发生构象变化。我们还在用核磁共振研究这种构象变化的动力学。我们需要收集的核磁共振实验包括：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。将测量处于各向同性和部分排列状态的样品的剩余偶极耦合(通过添加PF1噬菌体)。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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).