Dynamics of RNA-Peptide Complex Formation

关于RNA-多肽复合体的结构，有大量的核磁共振和X射线数据。 然而，关于它们形成的热力学和动力学方面的信息很少。我们建议使用双色荧光相关光谱(DcFCS)和单对荧光共振能量转移(SpFRET)来研究与RNA构建的多肽形成相关的动力学和热力学。本研究将重点研究牛免疫缺陷病毒反式活化区(BIV TAR)、BIV反式活化肽(BIV TAT)、人类免疫缺陷病毒1型Rev反应元件RNA构建(HIV-1 RRE)、HIV-1病毒表达调控肽(HIV-1 Rev)及其各自的复合体。我们的具体目标是： (1)分析BIV Tat多肽-BIV TAR RNA复合体的形成。 (A)复杂地层的热力学(三角洲H、三角洲G、三角洲)。 (B)改性络合物形成的热力学(增量H、增量G、增量)。 (C)BIV TAT多肽β发夹形成的动力学。 (D)BIV TAR RNA沟槽的形成动力学。 (2)分析HIV-1 Rev多肽-RNA复合体的形成。 (A)HIV-1 Rev肽-RNA复合体形成的热力学(DeltaH、DeltaG、Deltaas)。 (B)HIV-1 Rev多肽-RNA复合体的动力学。 (C)使用和不使用REV肽时HIV-1 RRE构象的动态变化。 本研究的结果将有助于理解RNA与多肽结合的动力学过程。

There is a substantial amount of NMR and x-ray data on the structure of RNA-peptide complexes. However, there is little information about the thermodynamics and dynamics of their formation. We propose to study the dynamics and thermodynamics associated with the formation of RNA construct peptide complexes using dual color fluorescence correlation spectroscopy (DCFCS) and single pair fluorescence resonance energy transfer (spFRET). This research will focus on bovine immunodeficiency virus trans-activating region (BIV TAR), BIV trans-activator peptide (BIV Tat), human immunodeficiency virus type 1 rev response element RNA construct (HIV-1 RRE), HIV-1 regulator of viral expression peptide (HIV-1 Rev) and their respective complexes. Our specific aims are: (1) Analyze the formation of BIV Tat peptide-BIV TAR RNA complexes. (a) Thermodynamics (deltaH, deltaG, deltaS) of complex formation. (b) Thermodynamics (deltaH, deltaG, deltaS) of modified complex formation. (c) Dynamics of BIV Tat peptide beta hairpin formation. (d) Dynamics of BIV TAR RNA groove formation. (2) Analyze the formation of HIV-1 Rev peptide - RNA complexes. (a) Thermodynamics (deltaH, deltaG, deltaS) of HIV-1 Rev peptide-RNA complex formation. (b) Dynamics of HIV-1 Rev peptide - RNA complexes. (c) Dynamics of HIV-1 RRE conformational changes with and without Rev peptide. The results of this study will contribute to understanding the dynamical process of RNA-peptide binding.