DYNAMIC MODES OF SPIN LABELED RNA

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 HIV病毒体生命周期中的蛋白质/RNA相互作用涉及异质环境中的复杂相互作用。电子自旋共振（ESR）是研究蛋白质/RNA相互作用的有力实验工具，当相互作用物种具有有限的内在结构时。多频ESR对微秒至纳秒时间尺度上的旋转动力学细节特别敏感，其中发生许多生化重要过程。在许多以前的研究和仔细的数值算法开发的基础上，它是已知的如何提取可靠的，翔实的结果，从多频ESR谱。我们建议研究自旋标记的TAR-RNA与蛋白质NCp 7相互作用的模型系统，由于其灵活的结构，很难通过其他手段进行研究。特别是，我们打算使用多频ESR来研究动态和结构方面的发夹结构蛋白的折叠和识别。 在高盐环境中，在240 GHz下，自旋标记的TAR-RNA与相关的NCp 7已经获得了有希望的初步结果。

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. Protein/RNA interactions in the life cycle of the HIV virion involve complex interactions in a heterogeneous environment. Electron Spin Resonance (ESR) is a powerful experimental tool to study protein/RNA interactions when the interacting species have limited intrinsic structure. Multifrequency ESR is particularly sensitive to details of rotational dynamics on the microsecond to nanosecond timescale, where many biochemically significant processes occur. On the basis of many previous studies and careful numerical algorithm development, it is known how to extract reliable, informative results from multifrequency ESR spectra. We propose to study a model system of spin-labeled TAR-RNA interacting with the protein NCp7 that would be difficult to study by other means due to its flexible structure. In particular, we intend to use multifrequency ESR to study dynamic and structural aspects of hairpin oligonucleotide-nucleocapsid protein folding and recognition. Promising initial results have been obtained at 240 GHz on spin-labeled TAR-RNA with associated NCp7 in a high salt environment.