Nano-lmaging APOBECS Interactions

APOBECSG（A3 G）和APOBECSF（A3 F）参与各种抗病毒蛋白和核酸的交易。重要的主要相互作用包括蛋白质间相互作用、结合单链核酸（RNA和ssDNA）以及HIV Vif的中和作用。这些分子间相互作用的充分阐明是必要的，以了解这些细胞蛋白抑制HIV复制的机制。寡聚化在其活性中的作用是一个非常重要的问题。这些是纳米级的相互作用，AFM是能够解决上述问题的首选方法。原子力显微镜目前达到的发展水平，使这一独特的仪器，以提供丰富的重要信息，在纳米尺度上的生物分子复合物的表征。分子系统的拓扑分析是AFM应用于表征蛋白质-DNA系统的结构和特异性的最初领域。除了成像，AFM能够直接测量系统内的分子相互作用，这十年的研究导致了AFM应用在这一领域的巨大进步。我们的初步研究已经使我们能够提供A3 G与ssDNA底物结合的第一个图像。我们进一步预计，使用所有形式的AFM技术将有助于我们详细说明这些复合物，剖析蛋白质和化学要求，并将关键观察扩展到ASF。为了帮助实现这些总体目标，我们将追求以下三个具体目标：目标1。图像A3复合物在纳米级分辨率使用原子力显微镜（计划目标1和2）;目标2。使用力谱（程序目标1和2）和目标3测量A3-ssDNA复合物内的相互作用。使用AFM成像和探测方式表征A3-Vif相互作用（计划目标3）。该项目下的研究是我们整体计划项目的一个组成部分，旨在全面了解A3-AS 3 A3-核酸和A3-Vif相互作用。 纳米级研究将为评估任何未来通过利用AS-Vif相互作用发挥作用的疗法提供平台。

APOBECSG (A3G) and APOBECSF (A3F) are involved in various anti-viral protein and nucleic acid transactions. Important primary interactions include interprotein interactions, binding single-stranded nucleic acids (RNA and ssDNA), and neutralization by HIV Vif. The full elucidation of these intermolecular interactions is necessary to understand the mechanism by which these cellular proteins inhibit HIV replication. The role of oligomerization in their activity is an issue of great importance. These are interactions at the nanoscale level, and AFM is the method of choice capable to addressing problems listed above. AFM currently reached a level of development that enables this unique instrument to provide a wealth of important information to the characterization of biomolecular complexes at nanoscale. The topographic analysis of molecular system is the initial area of the AFM application for characterization of the structure and specificity of protein-DNA systems of a broad complexity. In addition to imaging, AFM is capable of direct measurements of molecular interaction within the system and research during this decade led to a dramatic progress in this area of the AFM applications. Our Preliminary Studies have already enabled us to provide the first images of A3G bound to ssDNA substrates. We further anticipate that the use of all modalities of AFM technology will help us detail these complexes, dissect the protein and chemical requirements, and extend key observations to ASF. To help achieve these overarching objectives, we will pursue the following three Specific Aims: Aim 1. Image A3 complexes at nanoscale resolution using AFM (Program Objectives 1 & 2); Aim 2. Measure interactions within A3-ssDNA complexes using force spectroscopy (Program Objectives 1 & 2) and Aim 3. Characterize A3-Vif interactions using AFM imaging and probing modalities (Program Objectives 3). The studies under this project are an integral component of our overall Program Project aiming to provide a comprehensive understanding of A3-AS3 A3-nucleic acid, and A3-Vif interactions. The nanoscale studies will provide a platform for evaluating any future therapeutics that function by leveraging the AS-Vif interaction.