TETRALOOP RECEPTOR RNA STRUCTURAL INVESTIGATION

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 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，介导大的非编码RNA的三级折叠。 我们正在研究的四环受体结构作为温度和离子条件的函数。 以前的工作表明，四环受体经历了一个构象变化后，四环结合。 我们也正在研究这种构象变化的动力学NMR。 我们需要收集的NMR实验包括：1D（1H），2D NOESY，2D 1H-15 N HSQC（正常和TROSY增强），2D 1H-15 N HNN COSY，2D 1H-13 C HSQC（正常和TROSY增强），2D NOESY，2D 1H，13 C过滤/编辑NOESY，3D 1H（13 C）1H NOESY-HSQC，3D 1H（13 C）1H HCCH TOCSY和3D 1H（13 C）1H HCCH COSY。将测量各向同性和部分对齐状态下样品的残留偶极耦合（通过添加Pf 1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).