STRUCTURAL DETERMINATION OF RNA FOLDING INTERMEDIATES

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。 我们的目标是研究冷冻电子显微镜(Cryo-EM)和单粒子重建(SPR)在小RNA结构和折叠研究中的适用性。这些RNA的大小至少比这项技术通常设想的极限小两倍。我们的初步结果表明，冷冻-EM可以直接成像不同结构和功能状态的RNA的构象。这种方法对于阐明RNA的结构和功能非常有用，特别是那些通过构象调节和/或折叠过程来开启/关闭功能的RNA，例如催化化学反应的核酶或调节基因表达的核糖开关。低温电磁/表面等离子体共振技术需要广泛的计算能力。我们一直在使用需要几天时间才能处理单个数据集的计算机。TeraGrid提供的计算资源将大大加强这项研究。

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 aim to investigate the applicability of cryo-electron microscopy (cryo-EM) and single particle reconstruction (SPR) to structural and folding studies of small RNAs. These RNAs are at least two-times smaller in size than the generally conceived limits for this technique. Our preliminary results indicate that cryo-EM can directly image conformations of RNAs in different structural and functional states. This approach is very useful for elucidating RNA structure and function, particularly those whose function is toggled on/off through conformational adjustments and/or folding proceses, e.g. ribozymes that catalyze chemical reactions or riboswitches that regulate gene expression. The cryo-EM/SPR technique requires extensive computational power. We have been working with computers that take days for the processing of a single data set. Computational resources provided by the Teragrid will significantly enhance this study.